Integrative fungal solutions for protecting bees

ABSTRACT

The present invention is based on a plurality of benefits from the extracts of mycelia of individual fungal species, and mixtures of species, to provide an armamentarium of defenses from multiple stressors in order to help bees survive a complex of symptoms collectively called colony collapse disorder (CCD). More particularly, the present invention utilizes specific concentrations of extracts from pure cultured mycelium from mushroom forming fungi to reduce harmful viruses in bees and to increase the longevity of bees.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/332,803, filed Oct. 24, 2016, which is a continuation-in-part of U.S.patent application Ser. No. 14/641,432, filed on Mar. 8, 2015, whichclaims priority to U.S. provisional patent application Ser. No.62/074,023, filed on Nov. 2, 2014, and which is a continuation-in-partof U.S. patent application Ser. No. 14/247,207, filed on Apr. 7, 2014,which claims priority to U.S. provisional patent application No.61/967,117, filed on Mar. 10, 2014, each of which are herebyincorporated by reference in their entirety.

BACKGROUND

The present invention relates to compositions containing extracts ofmycelia of fungal species, and their mixtures, to provide anarmamentarium of defenses from multiple stressors in order to help beessurvive a complex of symptoms collectively called colony collapsedisorder (CCD). More particularly the present invention utilizesspecific concentrations of consumable extracts from pure culturedmycelium from mushroom forming fungi to reduce harmful viruses in beesand to increase the longevity of bees.

Approximately 100,000 species of insects, birds and mammals are involvedin the pollination of flowering plants. This includes almost 20,000known species of bees (bees are members of the superfamily Apoidea,considered to be a clade Anthophila). The Food and AgricultureOrganization of the United Nations estimates that of the slightly morethan 100 crop species that provide 90 percent of food supplies for 146countries, 71 are bee-pollinated (mainly by wild bees), and severalothers are pollinated by thrips, wasps, flies, beetles, moths and otherinsects. The annual monetary value of pollination services in globalagriculture could be as high as $200 billion. Protecting thePollinators, Food and Agriculture Organization of the United Nations,December 2005. The co-evolution of plants and bees (Apis species) isfundamental to their mutual survival. The bees spread pollen and manyplants produce rich nectar in return.

Approximately 4,000 bee species are native to North America. With theintroduction of European (or “western”) honey bees (Apis mellifera) toNorth America by colonists, commercial orchards and farms that would notnormally be able to survive have thrived, although many New World cropsand native flowering plants are primarily dependent upon native beespecies for pollination. Asian agriculture is similarly dependent uponthe Asian (or “eastern”) honey bee (Apis cerana), although typically ona smaller and more regionalized scale (A. mellifera has also beenintroduced). Throughout agriculture the number of fruit, nut andvegetable crops benefitting from bee pollination is staggering, as arethe number of flowering trees, shrubs, and wildflowers. Indeed, it isdifficult to overstate the role of bees in the commercial production offood. The loss of bees we are experiencing now is unprecedented and ahuge threat to food security worldwide. In some regions of China, forinstance, the loss of bees has necessitated hand pollination to savecrops, a dauntingly difficult task.

A honey bee hive is a warm, moist, densely populated environmentinhabited by closely related individuals—the perfect setting forviruses, bacteria, fungi, protozoa, and mites. Bees have successfullyprotected themselves for millions of years from such threats with uniquecolony-level and individual-level host defense systems and immuneresponses, but these defenses may be breaking down as the result ofintense domestication of the European honey bee and multiple threats,including new anthropogenic stressors, resulting in a precipitousdecline in the number of feral honey bees and native bees in areasincluding North America, Europe and China from 1972 to 2006, and theemergence of Colony Collapse Disorder (“CCD”) in honey bees in 2006.

The domestic honey bee industry is dependent upon queen breeding, theprocess of selection that brings about the lines to be propagated, andqueen rearing, the process of producing and culling queen honey bees.The large majority of bee breeding in the United States is carried outby 10-15 large queen-producing companies, who exchange geneticinformation from about 500 breeder queens. Such limited geneticdiversity may contribute to susceptibility to various diseases, pests,or colony collapse disorder. Particularly damaging to the rearing ofqueens are viruses, especially the Black Queen Cell Virus and otherviruses including the Deformed Wing Virus, the Israeli Acute BeeParalysis Virus, and nearly two dozen others. More viruses areanticipated to be discovered that contribute to illness in bees,including queens, their brood, in workers, nurse bees and drones.

Colony losses and bee disappearances have occurred throughout thehistory of beekeeping (“apiculture”), including various honey beesyndromes in the 1880s, the 1900s through the 1920s, the 1960s and the1990s, such as “disappearing disease,” “spring dwindle,” “fall dwindle,”“autumn collapse” and “mystery disease.” In 2006, some beekeepers beganreporting unusually high losses of 30-90 percent of their hives. Thisdisappearing bee affliction was renamed “colony collapse disorder” (CCD,sometimes referred to as spontaneous hive collapse or Mary Celestesyndrome in the UK). CCD may or may not be related to the prior colonyloss syndromes; it may be a genuinely new disorder or a known disorderthat previously only had a minor impact.

CCD is now approaching 40% with many beekeepers; with the ‘factoryfarms,’ where up to 84,000 beehives are kept in one location, CCD canclaim more than 60%. This has raised the costs for almond treepollination, for example, from $25-30 per bee colony per ½ to 1 acre ofalmond orchard for 3 weeks to more than $250. More than ⅓ of all thenon-animal food Americans consume is dependent upon pollination frombees. Should this upward trend in bee colony losses continue, theeconomic and societal expenses could run into the hundreds of billionsof dollars.

The loss of the services provided by bees has other far-reachingimplications. For example, neem trees, the source of thousands ofpopular health, beauty, and insecticide products, are dependent uponpollination from bees. Interestingly, neem products that contain theactive ingredient, azadirachtin, are useful for limiting or killingmites, including Varroa mites that transmit diseases to bees, andincluding mites that transmit diseases to other animals and plants.Should bees be lost, so too will this vast resource of health productsand a natural insecticide.

The main symptoms of CCD are the disappearance of the worker class(resulting in very few or no adult “worker” bees in the hive), a livequeen and few to no dead bees on the ground around the colony. Oftenthere is still honey in the hive, immature capped brood bees are present(bees will not normally abandon a hive until the capped brood have allhatched) and the hive contains honey and bee pollen that was notimmediately robbed by neighboring bees. The hive is also slow to berobbed by colony pests such as wax moths or small hive beetles. Varroamites, a virus-transmitting parasite of honey bees, have frequently beenfound in hives hit by CCD. Collapsing colonies typically do not haveenough bees to maintain colony brood and have workers that consist ofyounger adult bees; the progression of symptoms may be rapid or slow (upto two years). The colony may have ample food stores and be reluctant toeat food provided by the beekeeper. See, for example, Honey Bees andColony Collapse Disorder, United States Department of AgricultureAgricultural Research Service (2013).

The reasons for increasing colony collapse are complex and appear to bethe result of multiple factors. Suggested causes include increasingurbanization and loss of biodiversity, particularly loss of wildflowermeadows and “weeds” that provided high quality bee forage, poornutrition and malnutrition, immunodeficiencies, microbial pathogensincluding viruses, bacteria, fungi and protozoa, both lethal andsub-lethal exposure to insecticides, fungicides and herbicides,beekeeper applied miticides and antibiotics, parasitic mites (Varroadestructor and V. jacobsoni mites and Acarapis woodi tracheal mites),the fungi Nosema ceranae and N. apis, heavy metals, toxic pollutants,natural plant toxins, biting insects, selective breeding in apicultureand loss of genetic diversity, climate change, concentrations of hives,and increased environmental stresses from drought and cold snaps, andcombinations of these factors. Another factor is the new nature of thebee business and changing beekeeping practices. In the USA, there arefew or, in many regions, no feral bees and domesticated bee colonies areoften trucked hundreds of miles from factory bee ‘livestock’ apiaries,conferring additional stress factors to colony health and facilitatingwider spread of infections and parasites amongst bee populations.

Although the exact cause(s) and mechanisms of CCD remain to beelucidated, it appears the combination of stressors is of importance,particularly 1) microbial viral and fungal pathogens such as IsraeliAcute Paralysis Virus (“IAPV”), the Black Queen Cell Virus (“BQCV”) andDeformed Wing Virus (“DWV”) and Nosema (a pathogenic fungi); 2)parasitic mites (particularly Varroa mites); 3) pesticides at lethal orsub-lethal doses, including neonicotinoid insecticides (such asclothianidin, thiamethoxam, and imidacloprid) and beekeeper-appliedmiticides (“BAM”) and other environmental stressors; 4) the managementstressors of beekeeping including increasing viral exchange from truckedbees (particularly in the midwinter almond pollination migration toCalifornia), and 5) honey bee diets including use of honey substitutesand exposure to pollen of low nutritional value as opposed to nativediverse pollen and nectar of high nutritional value. Research suggeststhat honey bee diets, parasites, diseases, and multiple pesticidesinteract to have stronger negative effects on managed honey beecolonies, while nutritional limitation and exposure to sublethal dosesof pesticides, in particular, may alter susceptibility to or theseverity of bee parasites and pathogens. Pettis et al., Crop PollinationExposes Honey Bees to Pesticides Which Alters Their Susceptibility tothe Gut Pathogen Nosema ceranae, PLOS ONE, Published: Jul. 24, 2013.

Honey Bee Host Defense and Immune System

Colonies of bees may be infected by several species of parasites ordiseases at any time, but the colony-level and individual-level immunesystems generally deal with the infections (with the possible exceptionof parasitic Varroa destructor mites) provided that environmentalconditions are favorable. In the case of colony collapse, that normallyeffective immune function is clearly faltering. After the introductionof the parasitic, non-native Varroa destructor mite in 1987 to theUnited States, and its prolific spread throughout apiary populations,bees today face unprecedented threats from these virus-vectoringarthropods—fighting the viruses they introduce with immune systemsweakened from exposure to complex cocktails of xenobiotic toxins. Thisconvergence of stressors is a formula for disaster and is evolutionarilyunprecedented. Additional stressors are the loss of plant biodiversityas forests are cut, wood is removed, and monoculture factory farmsflatten the native landscapes. Bees, both domesticated and wild, ourgreatest pollinators, are under assault from multiple vectors. Beeextinctions have already been reported from some regions of China andare expected to occur with increasing frequency throughout the world.

Honey bees have numerous physical, chemical, and behavioral defenses atthe local population, colony hive, cell, and individual bee levels. Thefirst line of colony and individual defense is to avoid allowingparasites to gain a foothold—bees spend large amounts of energy oncooperative “social immunity” behaviors including grooming their bodysurfaces (both self auto-grooming and allo-grooming of a nestmate),cooperative hygienic behavior to detect and remove diseased brood andcorpses of adult bees from the hive, cleaning the inner surfaces of thenest cavity and sterilizing all surfaces with antimicrobial secretionsin their saliva (such as glucose oxidase), and utilizing (sometimescalled “stealing”) components of the plant immune system by gatheringthe highly antimicrobial resins found at leaf buds and wounds,incorporating them into propolis and using the propolis to form anantimicrobial barrier around for the colony, including heavy use at theentrance, coating inner surfaces of the cavity and face of the comb andsealing cracks and crevices.

Individual Systemic Immune Response

Insects possess innate immunity, which is characterized by non-specificimmune reactions against invading pathogens, while lacking the complex“adaptive” or “acquired” immunity such as formation of antibodiesspecific to new pathogens. The defense mechanism in insects consists ofcellular and humoral immunity. In the cellular defense mechanism,plasmocytes and granulocytes are the major haemocytes that react toforeign invaders either by phagocytosis and/or encapsulation. A hallmarkof the humoral reactions is the synthesis and secretion ofanti-microbial peptides (AMPs) that accumulate in the hemolymph againstinvading pathogenic bacteria. Yoshiyama, Innate immune system in thehoney bee, Honeybee Research Group, National Institute of Livestock andGrassland Science. This “induced” response of antimicrobial peptides canlast for weeks, and it appears these peptides can be passed to nestmatesto confer resistance prior to infection. Oliver, Sick Bees—Part 3: TheBee Immune System, American Bee Journal, October 2010.

The bee antiviral response is based upon RNA interference (RNAi). RNAi“silences” the expression of genes between the transcription of thegenetic code and its translation into functional proteins. MicroRNA(miRNA, small non-coding RNAs that function in networks ofprotein-coding genes and cell physiological processes viatranscriptional and post-transcriptional regulation of gene expression)and small interfering RNA (siRNA, short double-stranded fragments) bindto specific messenger RNA (mRNA) molecules and increase or decreasetheir activity, for example protein production or defending cellsagainst viral nucleotide sequences. The miRNAs are a well-conserved,evolutionarily ancient component of genetic regulation found in manyeukaryotic organisms.

RNAi is initiated by the enzyme Dicer, which cleaves longdouble-stranded (dsRNA) molecules into short double stranded fragmentsof siRNAs. Each siRNA is unwound into two single-stranded ssRNAs, thepassenger strand and the guide strand. The guide strand is incorporatedinto the RNA-induced silencing complex (RISC). After integration intothe RISC, siRNAs base-pair to their target mRNA and cleave it, therebypreventing it from being used as a translation template. When the dsRNAis exogenous (for example, coming from infection by a virus), the RNA isimported directly into the cytoplasm and cleaved to short fragments byDicer.

Bees possess more RNAi pathway components relative to flies and appearto more readily mount a systemic RNAi response than do flies. It followsthat bees should be quite capable of battling viruses and arguably otherpathogens through knockdowns based on double-stranded RNAs ofpathogen-expressed genes (Evans/Spivak 2009). Notably, this form ofresponse to viral attack provides a long-term memory similar to thatresulting from the antibodies produced in mammals. Oliver, SickBees—Part 4: Immune Response to Viruses, American Bee Journal, November2010.

Viruses, Nosema and Microbial Pathogens

Bees are host to at least 18 viruses, nearly all being single-strandedRNA viruses. Some are “emerging” pathogens, such as Deformed Wing Virusand Acute Bee Paralysis Virus, which were once considered to be“economically irrelevant” (Genersch 2010) and then, with the arrival ofVarroa as a vector, began to devastate colonies. Oliver, Sick Bees—Part4: Immune Response to Viruses, American Bee Journal, November 2010.

Viral diseases include Chronic Paralysis Virus (CPV), Acute BeeParalysis Virus (ABPV), Israeli acute paralysis virus (IAPV), KashmirBee Virus (KBV), Black Queen Cell Virus (BQCV), Cloudy Wing Virus (CWV),Sacbrood Virus (SBV), Deformed Wing Virus (DWV), Kakugo Virus,Invertebrate Iridescent Virus type 6 (IIV-6), Lake Sinai Viruses (LSV1and LSV2) and Tobacco Ringspot Virus (TRSV). Within these viruses aremany subtypes whose virulence towards bees is currently beinginvestigated. More pathogenic viruses will likely be discovered. Theco-occurrence of more than one internalized virus further challenges theimmunological health of bees. Hence, there is a need for advantageousremedies, which are non-toxic, yet active against more than one virus.

Bees are also vulnerable to pathogen host shifts. The tobacco ringspotvirus can replicate and produce virions in Apis mellifera honey bees,resulting in infections throughout the entire body, including extensiveinfection of the nervous system and likely impacts on colony survival.TRSV was also found in the gastric cecum of Varrroa mites, suggestingthat Varrroa mites may facilitate the spread of TRSV in bees whileavoiding systemic invasion. Li et al., Systemic Spread and Propagationof a Plant-Pathogenic Virus in European Honeybees, Apis mellifera, mBio5(1):e00898-13. The virus, first observed in infected tobacco, is spreadthrough infected pollen of numerous plant species including soy andnumerous crops, weeds, and ornamentals.

Nosema apis is a microsporidium, recently reclassified as a fungus,which invades the intestinal tracts of adult bees and causes Nosemadisease, also known as nosemosis. Nosema infection is also associatedwith Black Queen Cell Virus and Kashmir Bee Virus. Nosema ceranae isbecoming an increasing problem on both the Asian honey bee Apis ceranaand the western honey bee.

Some honey bee viruses (DWV and KBV) and the fungi Nosema ceranae areable to infect other species of bees and wasps, and possibly Varroa gutcells; honey bees are likely the source of the bumblebee pathogens.Fürst et al., Disease associations between honeybees and bumblebees as athreat to wild pollinators, Nature, Volume:506, 364-366, (2014). Thisnew bee-to-bee vector could be a tipping point, causing wide scalecollapse of many native bee species, with consequences well beyond ourcontrol, or imagination. From a historical and biological perspective,this is an ‘all hands on deck’ moment. What evolution has provided usover millions of years can be lost in decades due to the humaninterventions whose incentives are short term in view at the expense ofthe long term.

Bacterial diseases of bees include American foulbrood (AFB), caused byPaenibacillus larvae, and European foulbrood (EFB), caused by thebacterium Melissococcus plutonius. Fungal diseases include Chalkbrood,caused by Ascosphaera apis, and Stonebrood, a fungal disease caused byAspergillus fumigatus, Aspergillus flavus, and Aspergillus niger. New,as yet unidentified, fungal pathogens are expected to co-occur or becomea primary cause of bee diseases in the future as humans further alterthe natural environment and cause unintended consequences from the useof transgenic crops, more broadly known as GMOs—genetically modifiedorganisms. Such potential fungal pathogens include Candida,Cryptococcus, Coccidioides and other yeast-like organisms. And yet, manyof these so-called pathogens, especially, for instance, thepre-sporulating forms of entomopathogenic fungi, have properties thatcan confer benefits to insects, including bees, provided that theirendogenous toxins are eliminated, reduced or altered so to not harmbees, thereby reducing the threat to bees by disease-causing,disease-bearing or disease-spreading organisms.

All honey bees are infected by more than one species of bacteria,including beneficial endosymbionts that offer protection against yeasts,chalkbrood, and foulbrood. Apparently healthy bees may also be infectedby more than one species of virus. The dynamics of bee-bacteria,bee-virus and virus-virus interactions are complex and poorlyunderstood. Certain bee viruses may enhance the virulence of otherviruses while some bee viruses may competitively suppress thereplication of others. So too there are likely bacteria-to-bacteria,bacteriophage-to-bacteria, fungi-to-bacteria, and fungi-to-virusinterrelationships scientists have yet to discover. Many virulent beeviruses can exist in an “unapparent” infection—one can detect thepresence of the virus in bees, but there are no noticeable negativeeffects due to the infection. An infection by a second virus or otherstressor may cause a dormant virus to start replicating. A number ofresearchers have found that the mere action of a Varroa mite feedingupon a bee (which includes injection of immune suppressants by the mite)may induce or activate the replication of unapparent and normallynon-pathological virus infections. Studies of immune responses have alsoshown that mites and viruses could alter transcript levels ofimmunity-related genes in their corresponding hosts. It is common forcollapsing colonies to be simultaneously infected with three or fourviruses, Varrroa mites, Nosema (ceranae and especially apis), andtrypanosomes. See Oliver, Sick Bees—Part 3: The Bee Immune System,American Bee Journal, October 2010.

Crithidia bombi is a trypanosomatid protozoan bee parasite known to haveserious effects on bumblebees, particularly under starvation conditions.The related Crithidia mellificae may be contributing to mortality in thehoney bee. Ravoet et al., Comprehensive Bee Pathogen Screening inBelgium Reveals Crithidia mellificae as a New Contributory Factor toWinter Mortality (2013), PLoS ONE 8(8): e72443.

Varroa Mites and other Parasites

Varroa destructor and Varroa jacobsoni are parasitic mites that feed onthe bodily fluids of bee adults, pupae, and larvae. Acarapis woodi is atracheal mite that infests the airways of the honey bee. The Asianparasitic brood mites Tropilaelaps clareae and T. mercedesae areconsidered serious potential threats to honey bees, although they havenot been found in the United States or Canada to date.

The Asian honey bee Apis cerana is the natural host to the Varroajacobsoni mite and the parasite Nosema ceranae. Having co-evolved withthese parasites, A. cerana exhibits more careful grooming than A.mellifera, and thus has a more effective defense mechanism againstVarroa and Nosema, which are becoming increasingly serious pests of thewestern honey bee.

Varrroa mites breaching bees' hygienic, mechanical, and physiologicalbarriers to invasion have increasingly acted as a vector for viruses aswell as causing major stress to bees. Widespread colony losses have onlybeen reported from countries is which Varroa is a problem (Neumann2010). Colonies without mites may be virus free (Highfield 2009), but upto 100% of colonies with Varroa may be infected by one or more viruses,even if there are no apparent symptoms (Tentcheva 2004). Oliver, SickBees—Part 1, American Bee Journal, August 2010.

Varrroa mites have been found to be far more susceptible to acids thanare honey bees. Organic acids such as oxalic acid, formic acid andlactic acid can be used as “natural miticides” or means for killingmites in the hive, as they are all naturally found in honey. Othernaturally occurring miticides not typically found in honey, such asthymol or various essential oils, may also be utilized. Oxalic acid istypically mixed with distilled water to prevent the formation of salts,resulting in an acidic solution with pH often times <1. That the beescan tolerate such a low pH while mites cannot is significant. The oxalicacid will capture calcium and other minerals from the exoskeleton of themites to form oxalates. When direct contact of oxalic or formic acidwith the chitinous like exoskeleton of the mites pulls out calcium, theexoskeleton is weakened, thus making the mites susceptible to otherstressors, including but not limited to infection or toxin exposure fromentomopathogenic fungi.

Besides known colony insect pests, such as the greater and lesser waxmoths and the small hive beetle, the phorid fly, previously known toparasitize bumblebees, may be emerging as a threat to honey bees. Coreet al., A New Threat to Honey Bees, the Parasitic Phorid Fly Apocephalusborealis (2012), PLoS ONE 7(1): e29639; Ravoet, supra.

Pesticides

Pesticides cause multiple forms of stress to bees. Agricultural sprayingmay affect honey bees and large-scale spraying programs for mosquitoes,gypsy moths, spruce worms and other insect pests may cause direct orindirect bee kills including native bumblebees and solitary bees. Thereis also a shift in the types of pesticides applied—many, such asneonicotinoids, are less toxic to vertebrates and the necessity ofrepeated application is reduced, but they act systemically and areabsorbed and distributed throughout the plant upon seed or soiltreatment, including distribution to the pollen and nectar.

Sub-lethal pesticide exposure, including exposure to cholinergicneonicotinoid insecticides (nicotinic receptor agonists) and/orcholinergic organophosphate miticides (acetylcholinesterase inhibitors),has been found to alter bee activity, development, oviposition,behavior, offspring sex ratios, flight and mobility, navigation andorientation ability, feeding behavior, learning, memory and immunefunction, population dynamics and increase susceptibility to andmortality from diseases, including Nosema. See, for example, Pettis,Crop Pollination Exposes Honey Bees to Pesticides Which Alters TheirSusceptibility to the Gut Pathogen Nosema ceranae, supra at 1.Fungicides and miticides used by beekeepers can have a pronouncedability on bees' ability to withstand parasite infection. Pettis, supraat 4. Often bees are exposed to a variety of pesticides, which may haveinteractive effects. See, for example, Di Prisco et al., Neonicitinoidclothianidin adversely affects insect immunity and promotes replicationof a viral pathogen in honey bees, PNAS 110, no. 46, Nov. 12, 2013,18466-18471; Pettis et al., Crop Pollination Exposes Honey Bees toPesticides Which Alters Their Susceptibility to the Gut Pathogen Nosemaceranae, PLoS ONE (2013); Palmer et al., Cholinergic pesticides causemushroom body neuronal inactivation in honeybees, Nature Communications,4:1634, (2013); Williamson et al., Exposure to multiple cholinergicpesticides impairs olfactory learning and memory in honeybees, TheJournal of Experimental Biology 216, 1799-1807 (2013); Derecka et al.,Transient Exposure to Low Levels of Insecticide Affects MetabolicNetworks of Honeybee Larvae, PLoS ONE 8(7), e68191 (2013).

Exposure to fungicides also kills or reduces the beneficial fungi foundon pollen—the result likely being a higher incidence of disease in honeybees, including Nosema infections and chalkbrood (ironically, fungaldiseases). The bee genome has relatively few genes that are related todetoxification compared to solitary insects such as flies andmosquitoes. Some of the most marked differences between bees and otherinsects occur in three superfamilies encoding xenobiotic detoxifyingenzymes. Whereas most other insect genomes contain 80 or more cytochromeP450 (CYP) genes, A. mellifera has only 46 cytochrome P450 genes, whilsthumans host about 60 CYP genes. Honey bees have only about half as manyglutathione-S-transferases (GSTs) and carboxyl/cholinesterases (CCEs),compared to most insect genomes. This includes 10-fold or greatershortfalls in the Delta and Epsilon GSTs and CYP4 P450s, members ofwhich clades have been linked to insecticide resistance in otherspecies. Claudianos et al., A deficit of detoxification enzymes:pesticide sensitivity and environmental response in the honeybee, InsectMolecular Biology, 15(5), 615-636 (2006).

Whereas bees evolved to deal with plant phytochemicals and naturaltoxins, they now must additionally metabolize and detoxify anthropogenicinsecticides, miticides, herbicides, fungicides and environmentalpollutants, an unprecedented evolutionary challenge.

Management Stressors of Beekeeping

Use of honey or pollen substitutes (such as sugar syrup; high fructosecorn syrup; bee candy; “grease patties” containing grease, sugar andoptionally salt or essential oils; or “pollen patties” containing soy,yeast and nonfat dry milk, which may have added pollen, possibly fromareas contaminated with pesticides) may be a contributing factor todeclining bee populations and CCD for several reasons. Malnutrition islikely a major factor in declining bee populations. Synthesized beediets simply do not provide the nutritional value obtained by bees froma mixture of quality pollens. Although quality proteins, carbohydratesand vitamins can be provided to honey bees in the lab, we still cannotkeep them alive more than two months in confinement on our best diets.Garvey, About Bee Nutrition . . . , Posts Tagged: from the UC Apiariesnewsletter—The California Backyard Orchard.

Honey contains several substances that activate nutrient sensing,metabolic, detoxification and immune processes in the European honey beeApis mellifera, plus other chemicals useful to honey bee health. Theenzymes are found on the pollen walls of flowers and enter the honey bysticking to the bees' legs. Ingestion of tree resins, balsams, and treesaps via incorporation into propolis or bee glue is also known to reducebee susceptibility to both insecticides and microbial pathogens andup-regulate the transcription of the detoxification genes. Honeysubstitutes or pollen patties, which don't contain these chemicals, maytherefore contribute to colony collapse disorder. See Mao, Wenfru,Schuler, Mary A. and Berenbaum, May R., Honey constituents up-regulatedetoxification and immunity genes in the western honey bee Apismellifera, PNAS USA 110(22), 8842-8846 (2013). Mao et al. found thatconstituents in honey derived from pollen and tree exudates, includingp-coumaric acid (=4-hydroxycinnamic acid), pinocembrin, pinobanksin andpinobanksin 5-methyl ether, are strong inducers of cytochrome P450 genesdetoxification genes via a number of CYP6 and CYP9 family members.Massively parallel RNA sequencing and RNA-seq analysis revealed thatp-coumaric acid specifically up-regulates all classes of detoxificationgenes as well as select genes for antimicrobial peptides required fordefense against pesticides and pathogens.

Those species of honey bees that nest in tree cavities use propolis toseal cracks in the hive, as do bees in domestic hives, although feralhoney bees coat the entire inner surface of their nesting cavity,whereas domesticated honey bees lay down comparatively little resin inbeekeeping hives. The coating of propolis has been demonstrated toinhibit AFB (Antúnez 2008), fungi, and wax moth; Spivak has demonstratedthat propolis from some regions is effective against Varroa and isinvestigating its effect on viruses. Of great interest is the finding(Simone 2009) that the abundance of propolis appears to decrease thenecessary investment in immune function of bees—thus, the bee colony, byself-medicating with antimicrobial chemicals from plants, incurs less ofa metabolic cost in fighting pathogens. Oliver, Sick Bees—Part 3: TheBee Immune System, American Bee Journal, October 2010.

Bears, Mushrooms and Bees

The inventor noticed, on one of his many forays in the old growthforests of the Olympic Peninsula, a conifer tree scratched by a bear (aphotograph appears in the book he authored, Mycelium Running: HowMushrooms Can Help Save the World, 2005, pg. 70, figure 75. Ten SpeedPress, Berkeley). The research literature on the inter-relationshipsbetween bears and mushrooms stated that Fomitopsis species, brownrotting polypore wood conks, including the frequently seen Fomitopsispinicola and the rarely seen Fomitopsis officinalis, were the mostcommon fungal species to grow after bear scratchings in conifer forestsof the Pacific Northwest and elsewhere. Forest scientists showed thatwhen bears scratch a living tree, they leave an open wood, and theFomitopsis species opportunistically gain an entry site for infection.After a scratching, sugar-rich resin often beads out as droplets,attractive to bears and bees. Indeed, when the author returned a fewyears later to the same tree deep in the old growth forests along thesouth fork of the Hoh River, Olympic Peninsula of Washington State,Fomitopsis pinicola mushrooms were fruiting from the now-fallen tree.

“On young conifers, particularly Douglas-fir trees, bears will ripstrips of bark off with their teeth to reach insects or thesweet-tasting sap found inside. The bear's teeth leave long verticalgrooves in the sapwood and large strips of bark are found around thebases of trees they peel. These marks are typically made from April toJuly, but the results may be seen all year. This foraging activity iscommon in tree plantations where large stands of trees are similarlyaged and of a single species.” Link, Living with Wildlife: Black Bears,Washington State Dept. of Fish and Wildlife.

For this reason, a bounty was placed upon bears by forest stakeholderssince the bears were thought to reduce the profitability of forests fortimber. Tens of thousands of bears were killed by hunters hired by thetimber companies. In the 1990s, it was discovered that bears actuallybenefit the forests by bring sea minerals, particularly phosphorus andnitrogen, due to their foraging for salmon and trout in the riversadjacent to the forests. One reason the lowland old growth forests areso much larger than old growth forests several thousand feet up inelevation, above the limit of the migrating fish, is that bears broughtthe carcasses of fish onto shore, benefitting the adjacent trees. Humansare particularly adept at making decisions contrary to their long-termbest interests due to a fundamental misunderstanding about theinterconnectedness of nature.

In Stamets, Growing Gourmet and Medicinal Mushrooms, 1993, p. 42-43, thecurrent inventor stated “For 6 weeks one summer our bees attacked a KingStropharia bed, exposing the mycelium to the air, and suckled thesugar-rich cytoplasm from the wounds. A continuous convoy of bees couldbe traced, from morning to evening, from our beehives to the mushroompatch, until the bed of King Stropharia literally collapsed. When areport of this phenomenon was published in Harrowsmith Magazine (Ingle,1988), bee keepers across North America wrote me to explain that theyhad been long mystified by bees' attraction to sawdust piles.” Althoughit may not have been clear to one of ordinary skill in the art if thebees were attracted to the mycelium, the lignin within the sawdust orwood resins within the sawdust, the inventor concluded “Now it is clearthe bees were seeking the underlying sweet mushroom mycelium.”

An urgent solution is needed to the problems of declining bee health andcolony collapse disorder.

The present inventor sees the intersection and interplay of severalmycological methods and compositions as a possible integrated solutionto CCD. Each one of these elements may be sufficient to cause an effectleading to preventing or reducing CCD. As an integrated platform ofpartial solutions, the totality of these methods will achieve asynergistic benefit. More particularly, this invention focuses on theantiviral and longevity enhancing effects from extracts from purecultured mycelium, diluted to within specific ranges, which profferbenefits to bees.

The basis of these compositions and methods include the extracellularexudates and extracts made therefrom, of the pure cultured mycelium,prior to fruitbody formation, in the mushroom species of the Agaricales,Polyporales and Hymenochaetales in combination or independently.Miticides including oxalic acid, preconidial mycelium and extracts ofthe preconidial mycelium of entomopathogenic fungi may optionally beused to control mites and other bee and hive parasites. Mixtures ofthese extracts and bee products such as bee food or bee treatment spraysoffer multiple solutions to help prevent CCD or help bees overcome CCD.Sustainable solutions to problems plaguing bees will be derived frompromoting their natural defenses through habitat enhancement viabeneficial fungi, such as introducing mushroom forming fungi that haveantiviral properties to wood, causing rot, and ultimately moist nestingcavities that can be helpful to bees.

The inventor has isolated various strains of fungi, including Fomitopsisofficinalis, Fomitopsis pinicola, Ganoderma applanatum, Ganodermaannularis, Ganoderma lucidum, Ganoderma resinaceum, Inonotus obliquus,Irpex lacteus, Phellinus linteus, Piptoporus betulinus, Pleurotusostreatus, Polyporus umbellatus, Schizophyllum commune, and Trametesversicolor that have demonstrated superior antiviral, antibacterial,antifungal and antiprotozoal properties.

Without being bound to any theory, the inventor would hypothesize thatthese mushroom species are rich in compounds that up-regulate genes fordetoxification and defense against pollutants, pesticides, and pathogensin animals, including humans and bees. By repeatedly culturing andexpanding non-sporulating sectors of entomopathogenic fungi, theinventor also discovered that such “pre-sporulating” or “preconidial”mycelium and extracts of preconidial mycelium emit odors and fragrances(ranging from Metarhizium anisopliae and Aspergillus flavus“butterscotch” to Beauveria bassiana “vanilla cola” and “hard Christmascandy”) and tastes are attractive to animals including humans and bothnon-social and social insects, which offer advantages in control ofpests such as Varrroa mites.

The inventor now hypothesizes that the Fomitopsis colonization of thewood from bear foraging and the entry wound site (see above) would leadto the production of enzymes (laccases, lignin peroxidases, cellulases),ergosterols and other sterols, mycoflavonoids and especially arrays ofnutritious complex polysaccharides that would not only soften the wood,provide water, nutrition, and emit fragrances, all of which wouldattract bees, while the extracellular exudates being secreted by themycelium would be rich in p-coumaric acids and coumarins and theglycosides of unsubstituted and substituted benzoic, cinnamic andcoumaric acids, all stimulating the up-regulation of innate cytochromep450 genes and enzymes and also providing antiviral and antibacterialagents, all expressed during the decomposition of the infected tree. Acomplex fungal tree nectar is exuded, one that provides physiologicalbenefits and boosts the innate immunity of bees via numerous pathways asthe trees decompose. In some instances, bees nest within these logs orin the ground beneath them, benefitting from long-term contact. The beescan then incorporate these beneficial agents into their honey, propolisand combs so to as to protect the brood, the queen and ultimately thecolony. A plurality of virostatic molecules may result in a netreduction of viruses whereas the individual molecules may not.

The inventor also hypothesizes that combinations of the fungal speciesincluding but not limited to their resident phenols above and below willhave additive or even synergistic consequences, including regulation andup-regulation of nutrient-sensing, metabolic, detoxification, immunityand antimicrobial peptide genes and systems. This invention speaksdirectly to the link between the contact bees have with fungi that arebeneficial, not only nutritionally, but especially in activating thecytochrome P450 pathways for deactivating and metabolizing xenobioticand anthropogenic toxins.

The current invention provides a plurality of partial solutions toprovide scientists, farmers, biotechnologists, policy makers and thoughtleaders with biological tools of practical and scalable remedies beforeecological collapse forces us to ever-limiting options as biodiversityplummets. The combination of these partial solutions cumulatively andsynergistically provide that which is necessary for bees to overcomeCCD.

Extracts of Fomitopsis pinicola, Fomes fomentarius, Inonotus obliquus,Ganoderma lucidum, Ganoderma resinaceum (which is synonymous withGanoderma lucidum var. resinaceum) and Schizophyllum commune have nowbeen found to be effective in reducing the viral burden of honey beesand extending the life or worker bees.

“As an entomologist with 39 years of experience studying bees, I amunaware of any reports of materials that extend the life of worker beesmore than this.”*—Walter S. Sheppard, Ph.D., Chair, Dept. of Entomology,Washington State University (WSU). *Stamets et al., unpublished data, msin preparation.”

“I have never seen such strong antiviral activity against bee viruses asI have seen with Stamets' extracts.”*—Jay Evans, Ph.D., AgriculturalResearch Services, USDA. *Stamets et al., unpublished data, ms inpreparation.”

The inventor now anticipates, as a consequence of this invention, thatother woodland polypore mushrooms, for instance the birch polypore,Piptoporus betulinus, and numerous other woodland species will havegreater and lesser antiviral and longevity enhancing effects on beehealth when the extracts of the pure cultured mycelium are diluted towithin an optimal range, and presented as food, in the feed water, intohoney, pollen patties, propolis, or even sprayed onto bees orincorporated into the wood frames used to construct bee hives orincorporated into sticky strips applied to bee hives. The predominantviral species of concern are Deformed Wing Virus, Lake Sinai virus,Sacbrood virus, Israeli Acute Paralysis Virus and the Black Queen CellVirus, each one of which may exacerbate the activity of other viruses,and pathogens, as immunity fails from the deleterious cumulative effectfrom these and other multiple stressors.

As Albert Einstein noted, “We cannot solve our problems with the samethinking we used when we created them.” This patent follows thisphilosophy by offering a complex platform of synergistic solutionsaddressing a multiplicity of problems, which ultimately help beesovercome colony collapse disorder. With these hypotheses in mind, theinventor sees use of a wide array of Basidiomycetes, wood-decomposingfungi to develop a fungal bioshield, a “bee mycoshield” of protectionfrom the stressors leading to colony collapse disorder.

One embodiment described herein is a composition for use in improvingbee health comprising one or more bee feeding supplements and about 1%or less by volume of one or more aqueous ethanolic extracts of themycelium of Inonotus obliquus, Ganoderma resinaceum, Fomitopsispinicola, Fomes fomentarius, Schizophyllum commune, Trametes versicolor,Fomitopsis officinalis, Ganoderma applanatum, or combinations thereof.In one aspect the bee feeding supplements comprise one or more of water,sugars, sugar syrup, high fructose corn syrup water, bee candy, nectar,pollen, pollen patties, grease patties, propolis, bees wax, bee sprays,bee feed, protein supplements, or combinations thereof. In anotheraspect the composition improves bee health by increasing longevity bymore than about 1%. In another aspect the composition improves beehealth by increasing longevity by more than about 3%. In another aspectthe composition improves bee health by increasing longevity by more thanabout 5%. In another aspect the composition improves bee health byreducing viral load by more than about 1%. In another aspect thecomposition improves bee health by reducing viral load by more thanabout 15%. In another aspect the composition improves bee health byreducing viral load by more than about 25%. In another aspect thecomposition improves bee health by increasing longevity and reducingviral load by an LV index of more about than 1. In another aspect thecomposition improves bee health by increasing longevity and reducingviral load by an LV index of more than about 50. In another aspect thecomposition improves bee health by increasing longevity and reducingviral load by an LV index of more than about 200. In another aspect thebee feeding supplements and the aqueous ethanolic extracts are containedin a kit. In another aspect the composition further comprises one ormore second mycelium extracts from Antrodia cinnamonea, Ganoderma atrum,Ganoderma brownii, Ganoderma curtisii, Ganoderma lucidum, Ganodermalingzhi, Ganoderma oregonense, Ganoderma tsugae, Fomitopsis officinalis(Laricifomes officinalis), Fomitiporia robusta, Heterobasidion annosum,Inonotus hispidus, Inonotus andersonii, Inonotus dryadeus, Laetiporuscincinnatus, Laetiporus suiphureus, Laetiporus conifericola, Lenzitesbetulina, Phellinus igniarius, Phellinus linteus, Phellinus pini,Piptoporus betulinus, Polyporus elegans, Stereum complicatum, Stereumhirsutum, Stereum ostrea, Trametes elegans, Trametes gibbosa, Trameteshirsuta, Trametes villosa, Trametes cingulata, Trametes ochracea,Trametes pubescens, Trametes ectypa, Trametes aesculi, Wolfiporia cocos,Agaricus augustus, Agaricus blazei, Agaricus bonardii, Agaricusbrasiliensis, Agaricus campestris, Agaricus lilaceps, Agaricussubrufescens, Agaricus sylvicola, Agrocybe pediades, Agrocybe aegerita,Agrocybe arvalis, Agrocybe praecox, Clitocybe odora, Conocybe cyanopus,Conocybe lacteus, Conocybe rickenii, Conocybe smithii, Conocybe tenera,Coprinopsis nivea, Coprinopsis lagopus, Coprinus comatus, Coprinusmicaceus, Gymnopus hydrophilus, Gymnopus peronatus, Hypholoma aurantiaca(Leratiomyces ceres), Hypholoma capnoides, Hypholoma sublateritium,Hypsizygus marmoreus, Hypsizygus tessulatus, Hypsizygus ulmarius,Lentinus ponderosus, Lepiota procera (Macrolepiota procera), Lepiotarachodes (Chlorophyllum rachodes), Lepista nuda, Mycena alcalina, Mycenapura, Mycena aurantiidisca, Panellus serotinus, Panaelous foenisecii,Panaeolus subbalteatus, Pleurotus columbinus, Pleurotus ostreatus,Pleurotus cystidiosus, Pleurotus pulmonarius, Pleurotus sapidus,Pleurotus tuberregium, Panellus stipticus, Panellus serotinus, Pluteuscervinus, Psathyrella aquatica, Psathyrella candolleana, Psathyrellahydrophila, Psilocybe allenii, Psilocybe azurescens, Psilocybecaerulescens, Psilocybe coprophila, Psilocybe cubensis, Psilocybecyanescens, Psilocybe ovoideocystidiata, Psilocybe stuntzii, Psilocybesubaeruginosa, Stropharia aeruginosa, Stropharia cyanea, Strophariarugosoannulata, Stropharia semiglobata, Stropharia semiglboides,Stropharia squamosa, Stropharia thrausta, Stropharia umbonatescens,Termitomyces robusta, Volvariella bombycina, Volvariella volvacea orcombinations thereof and wherein the composition comprises a totalamount of about 1% or less by volume of mushroom mycelium extract.

Another embodiment described herein is a composition for use inimproving bee health comprising about 1% or less by volume of one ormore extracts from Inonotus obliquus, Ganoderma resinaceum, Fomitopsispinicola, Fames fomentarius, Schizophyllum commune, Trametes versicolor,Fomitopsis officinalis, Ganoderma applnatum, or combinations thereof; orabout 10% or less by volume of one or more extracts from Fomesfomentarius, Trametes versicolor, or combinations thereof; and one ormore bee feeding supplements; and an effective amount of a preservative.In one aspect the bee feeding supplement comprises one or more of water,sugars, sugar syrup, high fructose corn syrup water, bee candy, nectar,pollen, pollen patties, grease patties, propolis, bees wax, bee sprays,bee feed, protein supplements, or combinations thereof. In anotheraspect the mycelium is cultivated on a substrate comprising solidsubstrates or liquid substrates. In another aspect the preservativecomprises ethanol, isopropanol, methanol, butyl alcohol, other C₂-C₆alcohols, benzalkonium chloride, benzalkonium chloride solution,benzethonium chloride, benzoic acid, benzyl alcohol, butylparaben,cetylpyridinium chloride, chlorobutanol, chlorocresol, cresol,dehydroacetic acid, ethylparaben, methylparaben, methylparaben sodium,phenol, phenylethyl alcohol, phenylmercuric acetate, phenylmercuricnitrate, potassium benzoate, potassium sorbate, propylparaben,propylparaben sodium, sodium benzoate, sodium dehydroacetate, sodiumpropionate, sorbic acid, thimerosal, thymol, or combinations thereof. Inanother aspect the extracts comprise aqueous ethanol mycelium extracts;dried aqueous ethanol mycelium extracts; supernatant remaining afterprecipitation of an aqueous mycelium extract with ethanol; supernatantfrom aqueous ethanol mycelium extract having a portion of the solventremoved; supernatant from aqueous ethanol mycelium extract having thesolvent removed; supernatant from aqueous ethanol mycelium extracthaving a portion of solvent and all of the precipitate removed;supernatant from aqueous ethanol mycelium extract having both thesolvent and precipitate removed; non-aqueous and non-ethanolic solventmycelium extracts; dried non-aqueous and non-ethanolic solvent myceliumextracts; subcritical carbon dioxide mycelium extract supercriticalcarbon dioxide mycelium extract glycerol mycelium extracts; steamdistilled extracts; microwave-assisted extracts; or combinationsthereof. In another aspect the composition comprises one or more secondmycelium extracts from Antrodia cinnamonea, Ganoderma atrum, Ganodermabrownii, Ganoderma curtisii, Ganoderma lucidum, Ganoderma lingzhi,Ganoderma oregonense, Ganoderma tsugae, Fomitopsis officinalis(Laricifomes officinalis), Fomitiporia robusta, Heterobasidion annosum,Inonotus hispidus, Inonotus andersonii, Inonotus dryadeus, Laetiporuscincinnatus, Laetiporus sulphureus, Laetiporus conifericola, Lenzitesbetulina, Phellinus igniarius, Phellinus linteus, Phellinus pini,Piptoporus betulinus, Polyporus elegans, Stereum complicatum, Stereumhirsutum, Stereum ostrea, Trametes elegans, Trametes gibbosa, Trameteshirsuta, Trametes villosa, Trametes cingulata, Trametes ochracea,Trametes pubescens, Trametes ectypa, Trametes aesculi, Woffiporia cocos,Agaricus augustus, Agaricus blazei, Agaricus bonardii, Agaricusbrasiliensis, Agaricus campestris, Agaricus lilaceps, Agaricussubrufescens, Agaricus sylvicola, Agrocybe pediades, Agrocybe aegerita,Agrocybe arvalis, Agrocybe praecox, Clitocybe odora, Conocybe cyanopus,Conocybe lacteus, Conocybe rickenii, Conocybe smithii, Conocybe tenera,Coprinopsis nivea, Coprinopsis lagopus, Coprinus comatus, Coprinusmicaceus, Gymnopus hydrophilus, Gymnopus peronatus, Hypholoma aurantiaca(Leratiomyces ceres), Hypholoma capnoides, Hypholoma sublateritium,Hypsizygus marmoreus, Hypsizygus tessulatus, Hypsizygus ulmarius,Lentinus ponderosus, Lepiota procera (Macrolepiota procera), Lepiotarachodes (Chlorophyllum rachodes), Lepista nuda, Mycena alcalina, Mycenapura, Mycena aurantiidisca, Panellus serotinus, Panaeolus foenisecii,Panaeolus subbalteatus, Pleurotus columbinus, Pleurotus ostreatus,Pleurotus cystidiosus, Pleurotus pulmonarius, Pleurotus sapidus,Pleurotus tuberregium, Panellus stipticus, Panellus serotinus, Pluteuscervinus, Psathyrella aquatica, Psathyrella candolleana, Psathyrellahydrophila, Psilocybe allenii, Psilocybe azurescens, Psilocybecaerulescens, Psilocybe coprophila, Psilocybe cubensis, Psilocybecyanescens, Psilocybe ovoideocystidiata, Psilocybe stuntzii, Psilocybesubaeruginosa, Stropharia aeruginosa, Stropharia cyanea, Strophariarugosoannulata, Stropharia semiglobata, Stropharia semigloboides,Stropharia squamosa, Stropharia thrausta, Stropharia umbonatescens,Termitomyces robusta, Volvariella bombycina, Volvariella volvacea orcombinations thereof and wherein the composition comprises a totalamount of about 1% or less by volume of mushroom mycelium extract.

Another embodiment described herein is a bee treatment compositioncomprising: about 1% or less by volume of one or more mycelium extractsobtained from Inonotus obliquus, Ganoderma resinaceum, Fomitopsispinicola, Fomes fomentarius, Schizophyllum commune, Trametes versicolor,Fomitopsis officinalis, Ganoderma applanatum, or combinations thereof,at least one miticide comprising synthetic miticides, natural miticidesor combinations thereof, an effective amount of at least onepreservative; and at least one solvent. In one aspect the composition isaerosolized to treat bees, beehives, beehive components, bees wax, areassurrounding beehives, areas to be pollinated by bees or areas frequentedby wild bees. In another aspect the solvent comprises water, ethanol, awater ethanol mixture, 3-methoxy-3-methyl-1-Butanol (MMB), PEG-400,glycerol, propylene carbonate, or combinations thereof. In anotheraspect the natural miticide comprises Neem extracts, oxalic acid, formicacid, lactic acid, thymol, spores of entomopathogenic fungi pathogenicto mites, hyphae of entomopathogenic fungi pathogenic to mites,preconidial mycelium of entomopathogenic fungi pathogenic to mites,extracts of preconidial mycelium of entomopathogenic fungi pathogenic tomites, or combinations thereof. In another aspect the mycelium iscultivated on a substrate comprising solid substrates or liquidsubstrates.

Another embodiment described herein is a bee contacted with about 1% orless by volume of one or more extracts of the mycelium of Inonotusobliquus, Ganoderma resinaceum, Fomitopsis pinicola, Fomes fomentarius,Schizophyllum commune, Trametes versicolor, Fomitopsis officinalis,Ganoderma applanatum, or combinations thereof. In one aspect the bee iscontacted by oral feeding, topical application, or combinations thereof.In another aspect the extract is an aqueous ethanolic extract. Inanother aspect the extract further comprises one or more miticides. Inanother aspect the miticide comprises Neem extracts, oxalic acid, formicacid, lactic acid, thymol, spores of entomopathogenic fungi pathogenicto mites, hyphae of entomopathogenic fungi pathogenic to mites,preconidial mycelium of entomopathogenic fungi pathogenic to mites,extracts of preconidial mycelium of entomopathogenic fungi pathogenic tomites, or combinations thereof. In another aspect the extract furthercomprises one or more preservatives.

Another embodiment described herein is a composition comprising a beefeeding supplement and about 1% or less by volume of one or moreextracts of the mycelium of Inonotus obliquus, Ganoderma resinaceum,Fomitopsis pinicola, Fomes fomentarius, Schizophyllum commune, Trametesversicolor, Fomitopsis officinalis, Ganoderma applanatum, orcombinations thereof. In another aspect the composition comprises one ormore preservatives. In another aspect the preservative comprisesethanol, isopropanol, methanol, butyl alcohol, other C₂-C₆ alcohols,benzalkonium chloride, benzalkonium chloride solution, benzethoniumchloride, benzoic acid, benzyl alcohol, butylparaben, cetylpyridiniumchloride, chlorobutanol, chlorocresol, cresol, dehydroacetic acid,ethylparaben, methylparaben, methylparaben sodium, phenol, phenylethylalcohol, phenylmercuric acetate, phenylmercuric nitrate, potassiumbenzoate, potassium sorbate, propylparaben, propylparaben sodium, sodiumbenzoate, sodium dehydroacetate, sodium propionate, sorbic acid,thimerosal, thymol, or combinations thereof. In another aspect thepreservative comprises ethanol, isopropanol, methanol, butyl alcohol,other C₂-C₆ alcohols, benzalkonium chloride, benzalkonium chloridesolution, benzethonium chloride, benzoic acid, benzyl alcohol,butylparaben, cetylpyridinium chloride, chlorobutanol, chlorocresol,cresol, dehydroacetic acid, ethylparaben, methyl paraben, methylparabensodium, phenol, phenylethyl alcohol, phenylmercuric acetate,phenylmercuric nitrate, potassium benzoate, potassium sorbate,propylparaben, propylparaben sodium, sodium benzoate, sodiumdehydroacetate, sodium propionate, sorbic acid, thimerosal, thymol, orcombinations thereof.

Another embodiment described herein is a composition for improving beehealth comprising a bee feeding supplement and about 10% or less byvolume of one or more mycelium extracts of Fomes fomentarius, Trametesversicolor, or combinations thereof. In one aspect the compositionfurther comprises one or more preservatives. In another aspect thecomposition further comprises an effective amount of a preservative. Inanother aspect the preservative comprises ethanol, isopropanol,methanol, butyl alcohol, other C₂-C₆ alcohols, benzalkonium chloride,benzalkonium chloride solution, benzethonium chloride, benzoic acid,benzyl alcohol, butylparaben, cetylpyridini um chloride, chlorobutanol,chlorocresol, cresol, dehydroacetic acid, ethylparaben, methylparaben,methylparaben sodium, phenol, phenylethyl alcohol, phenylmercuricacetate, phenylmercuric nitrate, potassium benzoate, potassium sorbate,propylparaben, propylparaben sodium, sodium benzoate, sodiumdehydroacetate, sodium propionate, sorbic acid, thimerosal, thymol, orcombinations thereof. In another aspect the feeding supplement for beescomprises one or more of water, sugar water, sugar syrup, sugars, highfructose corn syrup water, bee candy, nectar, pollen, pollen patties,grease patties, propolis, bees wax, bee sprays, bee feed, proteinsupplements, or combinations thereof. In another aspect the mycelium iscultivated on a substrate comprising solid substrates or liquidsubstrates. In another aspect the composition for improving bee healthimproves the health of bees by increasing longevity by more than about1%. In another aspect the composition for improving bee health improvesthe health of bees by increasing longevity by more than about 3%. Inanother aspect the composition for improving bee health improves thehealth of bees by increasing longevity by more than about 5%. In anotheraspect the composition for improving bee health improves the health ofbees by reducing viral load by more than about 1%. In another aspect thecomposition for improving bee health improves the health of bees byreducing viral load by more than about 15%. In another aspect thecomposition for improving bee health improves the health of bees byreducing viral load by more than about 25%. In another aspect thecomposition for improving bee health improves the health of bees byincreasing longevity and reducing viral load by an LV index of morethan 1. In another aspect the composition for improving bee healthimproves the health of bees by increasing longevity and reducing viralload by an LV index of more than 50. In another aspect the compositionfor improving bee health improves the health of bees by increasinglongevity and reducing viral load by an LV index of more than 200. Inanother aspect the composition for improving bee health additionallycomprises a natural miticide, a synthetic miticide, or combinationsthereof, and wherein the natural miticide comprises one or more of Neemextracts, oxalic acid, formic acid, lactic acid, thymol, spores ofentomopathogenic fungi pathogenic to mites, hyphae of entomopathogenicfungi pathogenic to mites, preconidial mycelium of entomopathogenicfungi pathogenic to mites, extracts of preconidial mycelium ofentomopathogenic fungi pathogenic to mites, or combinations thereof. Inanother aspect the composition further comprises one or more secondmycelium extracts from Antrodia cinnamonea, Ganoderma atrum, Ganodermabrownii, Ganoderma curtisii, Ganoderma lucidum, Ganoderma lingzhi,Ganoderma oregonense, Ganoderma tsugae, Fomitopsis officinalis(Laricifomes officinalis), Fomitiporia robusta, Heterobasidion annosum,Inonotus hispidus, Inonotus andersonii, Inonotus dryadeus, Laetiporuscincinnatus, Laetiporus suiphureus, Laetiporus conifericola, Lenzitesbetulina, Phellinus igniarius, Phellinus linteus, Phellinus pini,Piptoporus betulinus, Polyporus elegans, Stereum complicatum, Stereumhirsutum, Stereum ostrea, Trametes elegans, Trametes gibbosa, Trameteshirsuta, Trametes villosa, Trametes cingulata, Trametes ochracea,Trametes pubescens, Trametes ectypa, Trametes aesculi, Wolfiporia cocos,Agaricus augustus, Agaricus blazei, Agaricus bonardii, Agaricusbrasiliensis, Agaricus campestris, Agaricus lilaceps, Agaricussubrufescens, Agaricus sylvicola, Agrocybe pediades, Agrocybe aegerita,Agrocybe arvalis, Agrocybe praecox, Clitocybe odora, Conocybe cyanopus,Conocybe lacteus, Conocybe rickenii, Conocybe smithii, Conocybe tenera,Coprinopsis nivea, Coprinopsis lagopus, Coprinus comatus, Coprinusmicaceus, Gymnopus hydrophilus, Gymnopus peronatus, Hypholoma aurantiaca(Leratiomyces ceres), Hypholoma capnoides, Hypholoma sublateritium,Hypsizygus marmoreus, Hypsizygus tessulatus, Hypsizygus ulmarius,Lentinus ponderosus, Lepiota procera (Macrolepiota procera), Lepiotarachodes (Chiorophyllum rachodes), Lepista nuda, Mycena alcalina, Mycenapura, Mycena aurantiidisca, Panellus serotinus, Panaeolus foenisecii,Panaeolus subbalteatus, Pleurotus columbinus, Pleurotus ostreatus,Pleurotus cystidiosus, Pleurotus pulmonarius, Pleurotus sapidus,Pleurotus tuberregium, Panellus stipticus, Panellus serotinus, Pluteuscervinus, Psathyrella aquatica, Psathyrella candolleana, Psathyrellahydrophila, Psilocybe allenii, Psilocybe azurescens, Psilocybecaerulescens, Psilocybe coprophila, Psilocybe cubensis, Psilocybecyanescens, Psilocybe ovoideocystidiata, Psilocybe stuntzii, Psilocybesubaeruginosa, Stropharia aeruginosa, Stropharia cyanea, Strophariarugosoannulata, Stropharia semiglobata, Stropharia semigloboides,Stropharia squamosa, Stropharia thrausta, Stropharia umbonatescens,Termitomyces robusta, Volvariella bombycina, Volvariella volvacea orcombinations thereof and wherein the composition comprises a totalamount of 1% or less by volume of mushroom mycelium extract.

Another embodiment described herein is a method for treating beescomprising contacting bees, beehives, beehive components, bees wax,areas surrounding beehives, areas to be pollinated by bees or areasfrequented by wild bees with the compositions described herein.

Another embodiment described herein is the product made by the processesdescribed herein.

Another embodiment described herein is a process for making acomposition for improving bee health comprising inoculating a substratewith Inonotus obliquus, Ganoderma resinaceum, Fomitopsis pinicola, Fomesfomentarius, Schizophyllum commune, Trametes versicolor, Fomitopsisofficinalis, Ganoderma applanatum to produce an inoculated substrate;cultivating the inoculated substrate to produce mycelium; extracting themycelium to produce a mycelium extract; adding one or more preservativesto the extract; and combining the extract with a bee feeding supplement.In one aspect the extract comprises one or more of: an aqueous ethanolmycelium extract; a dried aqueous ethanol mycelium extract; asupernatant remaining after precipitation of an aqueous ethanol myceliumextract; a supernatant from aqueous ethanol mycelium extract having aportion of the solvent removed; a supernatant from aqueous ethanolmycelium extract having the solvent removed; a supernatant from aqueousethanol mycelium extract having a portion of solvent and all of theprecipitate removed; a supernatant from aqueous ethanol mycelium extracthaving both the solvent and precipitate removed; a non-aqueous andnon-ethanolic solvent mycelium extract; a dried non-aqueous andnon-ethanolic solvent mycelium extract; a subcritical carbon dioxidemycelium extract a supercritical carbon dioxide mycelium extract aglycerol mycelium extract; a steam distilled extract; amicrowave-assisted extract; or a combination thereof. In another aspectthe preservative comprises ethanol, isopropanol, methanol, butylalcohol, other C₂-C₆ alcohols, benzalkonium chloride, benzalkoniumchloride solution, benzethonium chloride, benzoic acid, benzyl alcohol,butylparaben, cetylpyridinium chloride, chlorobutanol, chlorocresol,cresol, dehydroacetic acid, ethylparaben, methylparaben, methylparabensodium, phenol, phenylethyl alcohol, phenylmercuric acetate,phenylmercuric nitrate, potassium benzoate, potassium sorbate,propylparaben, propylparaben sodium, sodium benzoate, sodiumdehydroacetate, sodium propionate, sorbic acid, thimerosal, thymol, orcombinations thereof.

Another embodiment described herein is the product made by the processesdescribed herein.

Another embodiment described herein is a process for making acomposition for improving bee health comprising inoculating a substratewith Inonotus obliquus, Ganoderma resinaceum, Fomitopsis pinicola, Fomesfomentarius, Schizophyllum commune, Trametes versicolor, Fomitopsisofficinalis or Ganoderma applanatum to produce an inoculated substrate;cultivating the inoculated substrate to produce mycelium; extracting themycelium to produce a mycelium extract; adding a preservative to theextract; and combining with a miticide. In one aspect the extractcomprises one or more of: an aqueous ethanol mycelium extract; a driedaqueous ethanol mycelium extract; a supernatant remaining afterprecipitation of an aqueous ethanol mycelium extract; a supernatant fromaqueous ethanol mycelium extract having a portion of the solventremoved; a supernatant from aqueous ethanol mycelium extract having thesolvent removed; a supernatant from aqueous ethanol mycelium extracthaving a portion of solvent and all of the precipitate removed; asupernatant from aqueous ethanol mycelium extract having both thesolvent and precipitate removed; a non-aqueous and non-ethanolic solventmycelium extract; a dried non-aqueous and non-ethanolic solvent myceliumextract; a subcritical carbon dioxide mycelium extract a supercriticalcarbon dioxide mycelium extract a glycerol mycelium extract; a steamdistilled extract; a microwave-assisted extract; or a combinationthereof. In another aspect the preservative comprises ethanol,isopropanol, methanol, butyl alcohol, other C₂-C₆ alcohols, benzalkoniumchloride, benzalkonium chloride solution, benzethonium chloride, benzoicacid, benzyl alcohol, butylparaben, cetylpyridinium chloride,chlorobutanol, chlorocresol, cresol, dehydroacetic acid, ethylparaben,methylparaben, methylparaben sodium, phenol, phenylethyl alcohol,phenylmercuric acetate, phenylmercuric nitrate, potassium benzoate,potassium sorbate, propylparaben, propylparaben sodium, sodium benzoate,sodium dehydroacetate, sodium propionate, sorbic acid, thimerosal,thymol, or combinations thereof. In another aspect the miticidecomprises Neem extracts, oxalic acid, formic acid, lactic acid, thymol,spores of entomopathogenic fungi pathogenic to mites, hyphae ofentomopathogenic fungi pathogenic to mites, preconidial mycelium ofentomopathogenic fungi pathogenic to mites, extracts of preconidialmycelium of entomopathogenic fungi pathogenic to mites, or combinationsthereof. In another aspect, the process further comprises combining thecomposition with one or more bee feeding supplements. In another aspectthe bee feeding supplement comprises one or more of water, sugars, sugarsyrup, high fructose corn syrup water, bee candy, nectar, pollen, pollenpatties, grease patties, propolis, bees wax, bee sprays, bee feed,protein supplements, or combinations thereof.

Another embodiment described herein is the product made by the processesdescribed herein. Another embodiment described herein is a compositionfor use in spraying bees, beehives, beehive components or bees wax,areas surrounding beehives, areas to be pollinated by bees or areasfrequented by wild bees to improve bee health, the compositioncomprising about 1% or less by volume of one or more aqueous ethanolicmycelium extracts from Inonotus obliquus, Ganoderma resinaceum,Fomitopsis pinicola, Fomes fomentarius, Schizophyllum commune, Trametesversicolor, Fomitopsis officinalis, Ganoderma applanatum, orcombinations thereof; one or more miticides; and one or more solvents.In one aspect the extract comprises one or more of an aqueous ethanolmycelium extract; a dried aqueous ethanol mycelium extract; asupernatant remaining after precipitation of an aqueous ethanol myceliumextract; a supernatant from aqueous ethanol mycelium extract having aportion of the solvent removed; a supernatant from aqueous ethanolmycelium extract having the solvent removed; a supernatant from aqueousethanol mycelium extract having a portion of solvent and all of theprecipitate removed; a supernatant from aqueous ethanol mycelium extracthaving both the solvent and precipitate removed; a non-aqueous andnon-ethanolic solvent mycelium extract; a dried non-aqueous andnon-ethanolic solvent mycelium extract; a subcritical carbon dioxidemycelium extract a supercritical carbon dioxide mycelium extract aglycerol mycelium extract; a steam distilled extract; amicrowave-assisted extract; or a combination thereof. In another aspectthe miticide comprises Neem extracts, oxalic acid, formic acid, lacticacid, thymol, spores of entomopathogenic fungi pathogenic to mites,hyphae of entomopathogenic fungi pathogenic to mites, preconidialmycelium of entomopathogenic fungi pathogenic to mites, extracts ofpreconidial mycelium of entomopathogenic fungi pathogenic to mites, orcombinations thereof. In another aspect the solvent comprises water,ethanol, a water ethanol mixture, 3-methoxy-3-methyl-1-Butanol (MMB),PEG-400, glycerol, propylene carbonate, or combinations thereof.

Another embodiment described herein is a composition for use in sprayingbees, beehives, beehive components or bees wax to improve bee healthcomprising about 1% or less by volume of one or more mycelium extractsselected from the group consisting of Inonotus obliquus, Ganodermaresinaceum, Fomitopsis pinicola, Fomes fomentarius, Schizophyllumcommune, Trametes versicolor, Fomitopsis officinalis, Ganodermaapplanatum, or combinations thereof; one or more miticides, one or morepreservatives; and one or more solvents. In one aspect the extractcomprises an aqueous ethanol mycelium extract; a dried aqueous ethanolmycelium extract; a supernatant remaining after precipitation of anaqueous ethanol mycelium extract; a supernatant from aqueous ethanolmycelium extract having a portion of the solvent removed; a supernatantfrom aqueous ethanol mycelium extract having the solvent removed; asupernatant from aqueous ethanol mycelium extract having a portion ofsolvent and all of the precipitate removed; a supernatant from aqueousethanol mycelium extract having both the solvent and precipitateremoved; a non-aqueous and non-ethanolic solvent mycelium extract; adried non-aqueous and non-ethanolic solvent mycelium extract; asubcritical carbon dioxide mycelium extract a supercritical carbondioxide mycelium extract a glycerol mycelium extract; a steam distilledextract; a microwave-assisted extract; or a combination thereof. Inanother aspect the miticide comprises Neem extracts, oxalic acid, formicacid, lactic acid, thymol, spores of entomopathogenic fungi pathogenicto mites, hyphae of entomopathogenic fungi pathogenic to mites,preconidial mycelium of entomopathogenic fungi pathogenic to mites,extracts of preconidial mycelium of entomopathogenic fungi pathogenic tomites, or combinations thereof. In another aspect the preservativecomprises ethanol, isopropanol, methanol, butyl alcohol, other C₂-C₆alcohols, benzalkonium chloride, benzalkonium chloride solution,benzethonium chloride, benzoic acid, benzyl alcohol, butylparaben,cetylpyridinium chloride, chlorobutanol, chlorocresol, cresol,dehydroacetic acid, ethylparaben, methylparaben, methylparaben sodium,phenol, phenylethyl alcohol, phenylmercuric acetate, phenylmercuricnitrate, potassium benzoate, potassium sorbate, propylparaben,propylparaben sodium, sodium benzoate, sodium dehydroacetate, sodiumpropionate, sorbic acid, thimerosal, thymol, or combinations thereof. Inanother aspect the solvent comprises water, ethanol, a water ethanolmixture, 3-methoxy-3-methyl-1-Butanol (MMB), PEG-400, glycerol,propylene carbonate, or combinations thereof.

Another embodiment described herein is a bee, beehive, beehivecomponent, bees wax, areas surrounding beehives, areas to be pollinatedby bees or areas frequented by wild bees having been treated with acomposition comprising a mycelium extract of Inonotus obliquus,Ganoderma resinaceum, Fomitopsis pinicola, Fomes fomentarius,Schizophyllum commune, Trametes versicolor, Fomitopsis officinalis,Ganoderma applanatum, or combinations thereof. In one aspect thecomposition comprises a spray of about 1% or less mycelium extract ofInonotus obliquus, Ganoderma resinaceum, Fomitopsis pinicola, Fomesfomentarius, Schizophyllum commune, Trametes versicolor, Fomitopsisofficinalis, Ganoderma applanatum, or combinations thereof; or a sprayof about 10% or less mycelium extract of Fomes fomentarius, Trametesversicolor, or combinations thereof.

Another embodiment described herein is a process for making acomposition for use in improving bee health comprising extracting atleast one mycelium selected from the group consisting of Inonotusobliquus mycelium, Ganoderma resinaceum mycelium, Fomitopsis pinicolamycelium, Fomes fomentarius mycelium, Schizophyllum commune mycelium,Trametes versicolor mycelium, Fomitopsis officinalis mycelium, Ganodermaapplanatum mycelium, or combinations thereof; and combining the extractwith one or more bee feeding supplements. In one aspect the extractingcomprises an aqueous ethanol mycelium extract; a dried aqueous ethanolmycelium extract; a supernatant remaining after precipitation of anaqueous ethanol mycelium extract; a supernatant from aqueous ethanolmycelium extract having a portion of the solvent removed; a supernatantfrom aqueous ethanol mycelium extract having the solvent removed; asupernatant from aqueous ethanol mycelium extract having a portion ofsolvent and all of the precipitate removed; a supernatant from aqueousethanol mycelium extract having both the solvent and precipitateremoved; a non-aqueous and non-ethanolic solvent mycelium extract; adried non-aqueous and non-ethanolic solvent mycelium extract; asubcritical carbon dioxide mycelium extract a supercritical carbondioxide mycelium extract a glycerol mycelium extract; a steam distilledextract; a microwave-assisted extract; or a combination thereof.

Another embodiment described herein is the product made by the processesdescribed herein.

Another embodiment described herein is the process of combining thecomposition with a preservative.

Another embodiment described herein is the product made by the processesdescribed herein.

Another embodiment described herein is a process for making acomposition for improving bee health comprising inoculating a substratewith Inonotus obliquus, Ganoderma resinaceum, Fomitopsis pinicola, Fomesfomentarius, Schizophyllum commune, Trametes versicolor, Fomitopsisofficinalis, Ganoderma applanatum utilizing a means for inoculation toproduce an inoculated substrate; cultivating mycelium on the inoculatedsubstrate; extracting the mycelium to produce a mycelium extract; addinga preservative to the extract; and combining the extract with a beefeeding supplement.

Another embodiment described herein is the product made by the processesdescribed herein.

Another embodiment described herein is a process for improving animalhealth comprising (a) producing one or more extracts of a medicinalmushroom mycelium; (b) testing the extracts for activity against humanviruses to select active antiviral extracts or active antiviralmycelium; (c) combining the active antiviral extracts or activeantiviral mycelium, or combinations thereof, with an animal feed toproduce an antiviral food; and (d) feeding an animal the antiviral food.

Another embodiment described herein is the product made by the processesdescribed above.

Another embodiment described herein is a process further comprising: (e)testing the animal for virus loads, longevity, health benefits, orcombinations thereof; and (f) selecting an animal feed based on testingthe animal.

Another embodiment described herein is the product made by the processdescribed above.

Another embodiment described herein is a method for treating bees andimproving bee health comprising any of the methods described herein.

Another embodiment described herein is a composition for treating beesand improving bee health comprising any of the compositions describedherein.

BRIEF DESCRIPTION OF THE DRAWINGS

All figures refer to aqueous ethanolic extracts of mycelium grown ongrain. Sugar enriched water was made by adding 10 kg of sugar to 10 L, aweight to volume measurement (w/v) which is equivalent to a weight toweight (w/w) measurement of 10,000 grams sugar to 10,000 grams water.After this sugar enriched water is created, extracts are then addedusing a volume to volume (v/v) metric.

FIG. 1 is a line graph showing percent survival of bees over time whengiven extracts of the mycelium of Inonotus obliquus (0.1%, 1%, and 10%)with sugar water as compared to a control population fed sugar wateronly.

FIG. 2 is a line graph showing the percent survival of bees over timewhen given extracts of the mycelium of Ganoderma resinaceum (0.1%, 1%,and 10%) with sugar water as compared to a control population fed sugarwater only.

FIG. 3 is a line graph showing the percent survival of bees over timewhen given extracts of the mycelium of Fomitopsis pinicola (0.1%, 1%,and 10%) with sugar water as compared to a control population fed sugarwater only.

FIG. 4 is a line graph showing the percent survival of bees over timewhen given extracts of the mycelium of Fomes fomentarius (1%) with sugarwater as compared to a control population fed sugar water only.

FIG. 5 is a graph of Kaplan-Meier (product-limit) survival estimatesshowing the fraction of bees surviving over time when given extracts ofthe mycelium of Fomes fomentarius (0.1%, 1%, and 10%) with sugar wateras compared to a control population fed sugar water only.

FIG. 6 is a bar graph showing total virus particles in a controlpopulation and bees given extracts of the mycelium of Inonotus obliquus(0.1%, 1%, and 10%) with sugar water as compared to a control populationfed sugar water only at time zero, one week and two weeks.

FIG. 7 is a bar graph showing total virus particles in a controlpopulation fed sugar water only and bees given extracts of the myceliumof Ganoderma resinaceum (0.1%, 1%, and 10%) with sugar water as comparedto a control population at time zero, one week and two weeks.

FIG. 8 is a bar graph showing total virus particles in bees givenextracts of the mycelium of Fomitopsis pinicola (0.1%, 1%, and 10%) withsugar water as compared to a control population fed sugar water only attime zero and one week.

FIG. 9 is a bar graph showing total virus particles in and bees givenextracts of the mycelium of Schizophyllum commune (0.1%, 1%, and 10%)with sugar water as compared to a control population fed sugar wateronly at time zero and two weeks.

FIG. 10 is a line graph showing cycle threshold for Black Queen CellVirus over time in a control population and bees given extracts of themycelium of Inonotus obliquus (1%) and Ganoderma resinaceum (1%) withsugar water as compared to a control population fed sugar water only.

DETAILED DESCRIPTION

Bees are increasingly dealing with new anthropogenic stressors. Overhundreds of millions of years fungi have evolved to fight viruses,bacteria, and other fungi; have evolved to infect parasites, includinginsects; have evolved enzymes to break down toxins; and have evolvedsubstances to up-regulate such processes. This means they offer apotential nutraceutical treasure trove of compounds useful forprotecting bees and other pollinators from such threats, including aplurality of antiviral, antibacterial, antifungal, and antiprotozoalcompounds, and compounds useful for up regulating the digestive,detoxification, and immune systems of bees.

Without being held to any one theory, the inventor hypothesizes that thefungal mycelium extracts specifically modulate, induce and increase theexpression of detoxification and xenobiotic metabolizing genes,specifically to up-regulate all classes of detoxification genes,increase midgut metabolism of pesticides, function as a nutraceuticalregulating immune and detoxification processes, up-regulate immune,metabolic and nutrient pathways (lipid and glucose-metabolizingpathways) and up-regulate genes encoding antimicrobial peptides.Moreover, select fungal species support the microbiome of beneficialmicroorganisms in the digestion systems of bees, and their compatibilityis an important species-to-species bridge, matching beneficial woodrotting fungi to the beneficial microbes resident in the hindgut ofbees. The extracts of the present invention are expected to beprebiotics for the natural microbiome within the bee's digestive organsas well as to confer antiviral benefits, all of which contribute toextending longevity of bees and their colonies, and their collectivefunctionality. Additionally, since this inventor has found that extractsfrom the mycelia of certain polypore mushroom species have activityagainst some viruses and not others, the use of these extracts can shiftthe populations of viruses within the virome so bees can developpopulations of non-deleterious viruses that have favorably selected forby these extracts, and as a result these benign viruses can becomeresident within bees and prevent pathogenic viruses from emerging orspreading. This same principle—that the mycelial extracts both reducepathogens while promoting beneficial microbes—can also apply topromoting beneficial bacteria with the microbiomes of bees, as well asin the microbiomes other animals and plants.

Since bees are under assault from multiple pathogens—mites, viruses,microsporidia, protozoa, phorid flies and exposure to airbornepollutants—finding a robust broad-based platform of protection to helpbolster the host immune defense of bees is of paramount importance. Forexample, developing methods for creating compositions using theextracellular exudates of the mycelium of select species of fungi,including but not restricted to Stropharia rugosoannulata and othermembers of the Strophariaceae, Fomitopsis pinicola and other members ofthe Fomitopsidaceae and Metarhizium anisopliae and other members of theClavicipitaceae, can help prevent colony collapse disorder. Many otherspecies of basidiomycetes and ascomycetes are also expected to confersimilar benefits in the course of research into the benefits ofbee-beneficial exudates secreted by the laboratory grown, pure culturedmycelium.

With regard to fungal extracts, mycelial extracts are preferred to“mushroom extracts” because the hyphae produce extracellular exudatesthat are rich in accessible water, oils, polysaccharides, amino acids, Bvitamins, coumarins, p-coumaric acids, phenols and polyphenols, as wellas ergosterols, enzymes, acids, including fatty acids, antibacterialsand antivirals. The individual hyphal threads of the mycelium emitcomplex scents that volatilize into the air whereas the mushrooms tendto be nutritionally dense but do not have the extensive, exposedcellular surface area as the same mass of mycelium. The mushroomfruitbody is composed of cellularly compacted hyphae, laminatedtogether, so only a small fraction of the mycelial mass in the fruitbodyis exposed to the atmosphere. Hence the mushroom fruitbodies lack thefragrance attributes of the mycelium from which they form. Since theseextracellular exudates can readily dissolve into solution, theseexudates can be more usefully incorporated into amendments, such aspollen patties, sugar solutions or water, bee sprays or foliar plantsprays, and are better attractants to bees and other insects than themushroom fruitbodies. This is not currently obvious to those skilled inthe arts of mycology or entomology, whose focus has been more on thefruitbodies and spores from fruitbodies, rather than the mycelium.

Although bees may seek the sugar rich droplets exuding from the myceliumrotting wood, the extracts at 100% are far too potent and toxic in mostspecies in their natural form to be of benefit. Even at 10% of ourstandard 35% ETOH/H₂O extract (i.e., “1×”), a majority the testedextracts were toxic. Hence, if bees were to sip these droplets innature, they would likely sicken, prematurely die, and not reapbenefits. The inventor directed this research with his team,additionally led by Dr. Regan Nally, ably assisted by David Sumerlin,Henry Moershel, Bulmaro Solano, Dusty Yao, Morgan Wolff, Blake Westman,Alex Taylor and others, discovered that when the laboratory pure cultureextracts were highly diluted, to 10%, some toxicity remained for mostmushroom species but when further diluted to 1% or 0.1% or less,longevity substantially increased, especially in midlife, when theworkers are at the peak of their vigor and most productive in theirforaging and pollen acquisition. Similarly, when the extracts werediluted, antiviral benefits were seen at the same time longevityincreased in several fungal species tested. This is especially importantas the reduction of the pathogen payload has an overall net benefit tothe quality of the hive's overall health and performance. By combiningextracts optimized for antiviral activity with extracts optimized forlongevity, greater benefits than either are anticipated. Combined withlongevity benefits, the bees can be more productive as foragers, asnurses taking care of the brood, and as helpers for hygiene control,with less illness and better able to cope with exogenous stressors. Inessence, the services that bees provide internally within the hive, andexternally for the environment, are substantially augmented utilizingthe methods and compositions described within this invention.

As with botanicals, it is expected that fungal extracts may be moreeffective than single constituents or drugs. See, for example, Elfawalet al., Dried whole-plant Artemisia annua slows evolution of malariadrug resistance and overcomes resistance to artemisinin, PNAS USA112(3):821-6 (2015).

Only recently, research has discovered that the mycelium has more genesturned on than the mushrooms that ultimately are formed from it. As wasnoted by Li et al., 2013, “The protein-coding genes were expressedhigher in mycelia or primordial stages compared with those in thefruiting bodies.” Li et al., “Complete mitochondrial genome of themedicinal mushroom Ganoderma lucidum.” PLoS ONE 8(8):e72038 (2013).

Moreover, the network-like structure of the mycelium allows forepigenetic evolution of strains that can be evolved to emit substancestargeted specifically for the benefit of bees. Such improvements areanticipated by the inventor as a method for making strains andcompositions more attractive to bees and more appropriate for helpingbees overcome CCD.

In essence, the inventor has devised a novel nutraceutical which is richis a wide array of coumarins, coumaric acids, phenols and polyphenols;and anti-viral, anti-fungal, anti-bacterial and anti-protozoal agents,and a wide diversity of specialized metabolites such as antioxidants andantimutagens, which are generated as a result of mycelium digestinggrains or wood and are attractive to bees and supportive of their hostdefense against stressors and diseases. The extracts of mushrooms usedmedicinally for human health have an unexpected benefit for bee health,including lowering antiviral counts and extending bee lifespans. Indeed,the fungal contribution to propolis and honey, as well as to pollen,augments the immune systems of bees, and by extension to people, onspecific, fundamental, complex levels. The inventor notes that extractsof mycelium grown on grain inoculated wood are expected to contain morepolyphenols, coumarins and compounds that up-regulate detoxification andimmunity genes in the bees, as opposed to extracts of mycelium grown vialiquid fermentation. Since nature may require decades, even millennia,before new beneficial associations can be established, with bees unableto react quickly enough to the recent advent of new herbicides,pesticides, fungicides and miticides, we can jump start—jumping ahead ofevolution—this process by giving these beneficial fungal species aprimary role in the pathways of bee biology and biochemistry to bolstertheir host defenses and prevent CCD. The chemical composition of fungalmycelium is complex and variable within and among the various mushroomphyla, families and genera, traits that makes fungal extracts a gooddefense against rapidly evolving pests and pathogens.

The mycelium in many fungal species will not form sporulatingstructures, including but not limited to mushroom formation; such fungiare also preferred for studying their mycelial extracts for beeattractancy and health.

Extraction of pure culture, laboratory mycelium on sterilized substratesis substantially different than naturally occurring myceliumform—structurally, quantitatively, and qualitatively. Moreover, growingthe pure culture mycelium on rice, for instance, a non-native substrate,away from the numerous other co-occurring microbes resident on naturallydecomposing wood, produces an arguably different substance than exudatesfrom decomposing wood resplendent with myriads of other organisms (agram of rotting wood naturally hosts tens of thousands of othermicroorganisms, including bacteria protozoa, other fungi, co-inhabitingwith or upon the wood decomposing mycelium). Hence the exudates from theraw mycelium in nature, containing a plurality of organisms, isfundamentally different and are unlikely to benefit bees with the sameantiviral and longevity benefits seen with the specifically diluted,pure culture extracts made from mycelium as described in the currentinvention. In other words, the bees benefit from several alterations andmanipulations by the inventor outside of nature: the exudates from thepure culture mycelium must be highly diluted within discreteconcentrations to show benefits. After finding the initial extracts tobe toxic, most researchers would have abandoned this line of inquiry.Indeed, when the inventor proposed this idea to entomologists andmycologists skilled in the art, they deferred to engage with theinventor as they expected toxicity and did not want to harm bees.Initial results, ironically, confirmed their suspicions. Some even saidmy idea was “preposterous” and wanted nothing to do with it. Anature-based product such as an extract has markedly differentcharacteristics as compared to mushroom mycelium, an extract's closestnaturally occurring counterpart in its natural state. Mycelium issubject to death and resultant decay, spoilage and bacterial or fungalcontamination, and even an aqueous extract or hot water extract ofmycelium (“mycelium juice”) is subject to rapid souring, spoilage anddecay from bacteria or fungi. In tests by the inventor, dead myceliumvisually sours in 48-72 hours which means it sours with bacteriamicroscopically after 24 hours if kept at room temperature. Aqueousextracts rapidly sour, with bacterial contamination being noticeable tothe eye after 48 hours at 75° F. (24° C.). Bacterial contamination issimilarly noticeable after one month or less at 34° F. (1° C.). Unlikethe naturally occurring counterparts, aqueous ethanolic extract withsufficient alcohol to act as a preservative, or aqueous extract with aneffective amount of natural or non-natural synthetic preservative, or a“dried” extract from which the water, alcohol and/or other solvent hasbeen removed, are either slower to contaminate and decay or, ifsufficient alcohol or other preservative is present or the extract isdried, not subject to bacterial, fungal or other microbial contaminationand resulting souring or spoilage and decay for years. Alcohol “acts asa natural preservative, preventing the souring of the protein-richsaccharides from microorganisms. Stamets, MycoMedicinals, 2002 (3^(rd)ed.). Alcohol is not naturally produced by these mushroom species in anysignificant quantity (<1%). Clearly extracts have markedly differingproperties than naturally occurring living or dead mushroom mycelium.

Additionally, aqueous, aqueous ethanolic or dried, solvent free extractsare structurally and functionally different than mycelium in that,unlike mycelium, they are soluble in bee feeding supplements such assugar water honey and water, whereas mycelium contains a largepercentage of solid insoluble components which may clog typical beefeeding equipment such as drip feeders designed for liquids and may clogsprayers. Accordingly, isolated extract products' solubility alsoresults in a functional change in characteristics and propertiessufficient to show a marked difference from the naturally occurringcounterpart(s). Also, by precipitating out heavy molecular weight betaglucans by adding ethanol >20-25%, a further sub-fraction is made fromthe extracted mycelium and this is more perishable than beta glucanssince their molecules are more easily metabolized due, in part, to theirsmaller molecular masses. Hence the alcohol acts as a criticalpreservative but also serves additional purposes by excludingconfounding molecules—precipitating beta glucans—thought and taught bymost experts who focus on this fraction for medicinal benefits.

Applicant would further note that there is no “naturally occurringextract” of a preferred polypore or gilled mushrooms—there is thenaturally occurring non-sterile mushroom fruitbody and the non-sterilemycelium growing in or embedded in the wood or other substrate. Myceliumof polypore fungi do not grow naturally on rice or grains. In creatingextracts preserved in ethanol solutions of >25% from living or deadmycelium, the supernatant is used for this invention. In addition, theinventor has found that aqueous extracts of mycelium are inactive untilsolids are precipitated out with ethanol. Repeated DNA testing of >25%ethanol preserved supernatant reveals no identifiable DNA in theextracts used for helping bees fight viruses and extend longevity.Although new technologies may eventually evolve to amplify geneticresidues in the supernatant extracts, the current state of the arttechnology cannot. Extracts are not natural products as natural productsof fungi typically have identifiable DNA. Therefore, the claimedextracts of mycelium and the naturally occurring, living or deadnon-sterile mycelium and fruitbodies have markedly different functionaland structural characteristics and, obviously, differing properties inmany regards.

Combinations of extracts of a longevity extending mushroom such asFomitopsis pinicola combined with an antiviral species such asFomitopsis officinalis, Inonotus obliquus, Fomes fomentarius, Ganodermalucidum or Ganoderma resinaceum, offers unique benefits. Combinations ofa polypore mushroom like Fomitopsis officinalis (which grows only onfirs and larch trees) and Inonotus obliquus or Fomes fomentarius (whichonly grow on deciduous trees not firs or larch) is unique, as theycannot co-occur in nature. Hence extracts of these species are a uniquecombination of ingredients not found in nature. In fact, given thenumber of species of mushroom-forming fungi with antiviral and longevityextending properties, this inventor anticipates complex and uniquecombinations of fungal species, especially those not necessarilyassociated on the same host trees. The author envisions being able to‘dial in’ combinations of species customized to the viral, ecological,genomic, xenobiotic, and other stressors challenging beesgeographically. Disease vector algorithms tracking viral epidemics andpandemics would assist in designing these formulas so to optimizeformulas, collect data, and track improvements.

Experience with alcoholic beverages demonstrates that beers or wineswith less than 12% alcohol content will “go off” very soon after beingexposed to air, while a fortified wine such as a sherry with a 17.5%alcohol content will survive a year or more after being uncorked, andspirits (34%+) will evaporate but not contaminate or sour. For themajority of herbal tinctures, an alcohol content of 25% is the acceptedstandard. This compensates for the fact that plant constituentsdissolved into the menstruum will effectively reduce the proportion ofalcohol in the finished product to around 20%, at which level thetincture can be expected to have an acceptable shelf-life. Applicantwould note that dilution of one part 96% alcohol to three parts watergives 24% alcohol, which is normally referred to as “25% alcohol” in theindustry. With regard to aqueous ethanolic mushroom mycelium extracts,20%, 25% or greater alcohol content is an effective amount ofpreservative and therefore is typically preferred.

Viruses, Fungi, Bacteria and Protozoa

Bees infected by viruses can lose immune function, as well as theability to perform other metabolic functions, as a result of the viruses“hijacking” the ribosomal machinery to their benefit, chemicallyinterfering with the crucial phenoloxidase cascade, suppressing immuneresponses before they are initiated, manipulating the host's immunesignaling network, disabling the host's antimicrobial peptides,interfering with the RNAi response and/or creating “superantigens” thatcan overwhelm the host immune system and otherwise adversely affectingbee health.

The exclusive dependence of viruses on the host cellular machinery fortheir propagation and survival make them highly susceptible to thecharacteristics of the cellular environment like short RNA mediatedinterference. It also gives the virus an opportunity to fight and/ormodulate the host to suit its needs. Thus, the range of interactionspossible through miRNA-mRNA cross talk at the host-pathogen interface islarge. These interactions can be further fine-tuned in the host bychanges in gene expression, mutations, and polymorphisms. In thepathogen, the high rate of mutations adds to the complexity of theinteraction network. Viruses either produce micro-RNAs or target hostmicro-RNAs essential to the host immune system. Scaria et al.,Host-virus interaction: a new role for microRNAs, Retrovirology, 2006,3:68; Oliver, Sick Bees—Part 4: Immune Response to Viruses, American BeeJournal, November 2010.

Mushroom mycelium produces a wide array of compounds that can beanti-bacterial or anti-viral. U.S. Pat. No. 8,765,138 to the inventordiscloses the antiviral activity of Fomitopsis officinalis, whichincludes activity against avian flu viruses and herpes simplex I & II.Other viruses are anticipated to be sensitive to the antivirals beingcoded and expressed by the mycelium of Fomitopsis officinalis, andindeed many species in the polyporaceae and Basidiomycetes fungi. Themycelial extracts are active against numerous viruses that harm bees,particularly but not limited to BQCV (Black Cell Queen Virus), IAPV(Israeli Acute Paralysis Virus), DWV (Deformed Wing Virus), TRV (TobaccoRingspot Virus), and their relatives. The active ingredients limitingviruses within extracts are varied, but two groups are polyphenolsincluding coumarins and sterols including dehydrosulpherinic acids,eburicoic acids and related compounds. Synergistic benefits betweenthese polyphenols and sterols can further boost the host defense ofbees. These compounds are resident within the complexes that includefatty acids, lipids, and sterols. As such, many other active ingredientsrelated to fatty acids, lipids and sterols having antiviral propertiesare expected to be of bee benefit. Many of these aforementionedcompounds known as bioflavonoids, and the species that produce them, areof interest because some of these species produce mycelium with brightyellowish colors, which may also serve to attract bees. Very littlework, if any, has been done by mycologists to detect the “colors” ofmyceliated wood visible to bees but invisible, or nearly so, to thehuman eye, especially light reflected in the ultraviolet bands.

The inventor has also discovered the antibacterial properties ofFomitopsis officinalis mycelial extracts against staph, tuberculosis,and E. coli bacteria. This antibacterial activity is likely to confer anadditional layer of protection from diseases carried by other organisms.These extracts will similarly have a positive influence in limiting thedeleterious effects from known and yet undiscovered bacteria that areharmful to bees, animals, and plants. See U.S. patent application Ser.No. 13/998,914 and related applications above.

It is expected that medicinal mushroom species substances useful inhumans will similarly prove useful in up-regulating of immune genes andbenefitting the bee's immune system. Since many such genes areevolutionarily conserved or similar, it is expected that the extracts ofthe mycelium of such mushrooms will similarly be useful in up-regulatinggenes and systems in bees to degrade and deal with infections.

A preferred effective dose varies from species to species, in partbecause the extracts can be, in common with most medicines, medicinal atlow doses and toxic at high doses. In addition, some species such asFomitopsis officinalis may have both strong antiviral effects and alower toxic threshold as compared to other medicinal species. Ingeneral, for all medicinal mushroom species mentioned herein by thisinventor, preferred doses range from 0.0001% to 50%, with a morepreferred range of 0.001%-25% and a most preferred range of 0.01% to15%. With many of the polypore extracts in particular, the results ingeneral indicate that the extracts need to be diluted to 10% or less, 1%or less or 0.1% or less to confer antiviral and longevity benefits tobees. A preferred dose added to liquid or solid bee nutrients forFomitopsis officinalis would be from 0.0001%-0.1%; a preferred dose forTrametes versicolor or Fomes fomentarius and F. pinicola would be from0.1% to 10% based on results that show both improved longevity andimproved reduction in viral load at 10% concentrations. Except forTrametes versicolor and Fomes fomentarius, in general 10% concentrationsdid not help increase bee longevity. In general, 1% is a preferred dosefor both grain and sawdust ethanolic extracts. Consistently, higherconcentrations, above 10% had adverse effects on overall lifespans.

Medicinal mushrooms and the mycelium of medicinal mushrooms are definedas mushrooms and mycelium that support health and nutrition. In thecontext of bees, this includes mushrooms and preferred mycelia that havethe effect of increasing longevity, increasing foraging abilities,increasing resistance to disease, increasing ability to detoxifyanthropogenic toxins, increasing parasite resistance, possessingantiviral, antibacterial and/or antifungal activity, and increasingbees' ability to better withstand stressors associated with the complexcollectively called ‘colony collapse disorder.’

Useful and preferred fungal genera include, by way of example but not oflimitation: the gilled mushrooms (Agaricales) Agaricus, Agrocybe,Armillaria, Clitocybe, Collybia, Conocybe, Coprinus, Coprinopsis,Flammulina, Giganopanus, Gymnopilus, Hypholoma, Inocybe, Hypsizygus,Lentinula, Lentinus, Lenzites, Lepiota, Lepista, Lyophyllum, Macrocybe,Marasmius, Mycena, Omphalotus, Panellus, Panaeolus, Sarcomyxa, Pholiota,Pleurotus, Pluteus, Psathyrella, Psilocybe, Schizophyllum, Stropharia,Termitomyces, Tricholoma, Volvariella, etc.; the polypore mushrooms(Polyporaceae) Albatrellus, Antrodia, Bjerkandera, Bondarzewia,Bridgeoporus, Ceriporia, Coltricia, Coriolus, Daedalea, Dentocorticium,Echinodontium, Fistulina, Flavodon, Fomes, Fomitopsis, Fomitiporia,Ganoderma, Gloeophyllum, Grifola, Heterobasidion, Inonotus, Irpex,Laetiporus, Meripilus, Oligoporus, Oxyporus, Phaeolus, Phellinus,Piptoporus, Polyporus, Poria, Schizophyllum, Schizopora, Trametes,Wolfiporia; the toothed mushrooms Hericium, Sarcodon, Hydnum, Hydnellumetc.; Basidiomycetes such as Auricularia, Calvatia, Ceriporiopsis,Coniophora, Cyathus, Lycoperdon, Merulius, Phlebia, Serpula, Sparassisand Stereum; Ascomycetes such as Cordyceps, Ophiocordyceps, Morchella,Tuber, Peziza, etc.; ‘jelly fungi’ such as Tremella; the mycorrhizalmushrooms, fungi such as Phanerochaete (including those such as P.chrysosporium with an imperfect state and P. sordida).

Suitable fungal species and genera include by way of example only, butnot of limitation: Agaricus augustus, A. blazei, A. brasiliensis, A.brunnescens, A. campestris, A. lilaceps, A. placomyces, A. subrufescensand A. sylvicola, Acaulospora delicata; Agrocybe aegerita, A. praecoxand A. arvalis; Albatrellus hirtus and A. syringae; Alpova pachyphloeus;Amanita muscaria; Antrodia carbonica, A. cinnamomea and A. radiculosa;Armillaria bulbosa, A. gallica, A. matsutake, A. mellea and A.ponderosa; Astraeus hygrometricus; Athelia neuhoffii; Auriculariaauricula and A. polytricha; Bjerkandera adusta and B. adusta;Boletinellus merulioides; Boletus punctipes; Bondarzewia berkeleyi;Bridgeoporus nobilissimus; Calvatia gigantea; Cenococcum geophilum;Ceriporia purpurea; Ceriporiopsis subvermispora; Clitocybe odora,Collybia albuminosa and C. tuberosa; Coltricia perennis; Coniophoraputeana; Coprinus comatus, C. niveus and ‘Inky Caps’; Cordycepsbassiana, C. variabilis, C. fads, C. subsessilis, C. myrmecophila, C.sphecocephala, C. entomorrhiza, C. gracilis, C. militaris, C.washingtonensis, C. melolontha, C. ravenelii, C. unilateralis, C.clavulata and C. sinensis; Cyathus stercoreus; Daedalea quercina;Dentocorticium sulphurellum; Echinodontium tinctorium; Fistulinahepatica; Flammulina velutipes and F. populicola; Flavodon flavus; Fomesfomentarius, F. lignosus; Fomitopsis officinalis, Fomitopsis cana, F.subtropica and F. pinicola; G. resinaceum, annularis, G. australe, G.atrum, G. brownii, G. collosum, G. sinensis, G. lingzhi, G. curtisii, G.japonicum, G. lucidum, G. resinaceum, G. neo-japonicum, G. oregonense,G. sinense, G. tornatum and G. tsugae; Gigaspora gigantea, G. gilmorei,G. heterogama, G. margarita; Gliocladium virens; Gloeophyllum sepiarium;Glomus aggregatum, G. caledonia, G. clarus, G. fasciculatum, G.fasiculatus, G. lamellosum, G. macrocarpum and G. mosseae; Grifolafrondosa; Gymnopus dryophilus, Gymnopus peronatus, Hebelomaanthracophilum and H. crustuliniforme; Hericium abietis, H. coralloides,H. erinaceus and H. capnoides; Heterobasidion annosum; Hypholomacapnoides and H. sublateritium; Hypsizygus ulmarius and H. tessulatus(=H. marmoreus); Inonotus hispidus and I. obliquus; Irpex lacteus;Lactarius deliciosus; Laetiporus sulphureus (=Polyporus sulphureus), L.conifericola, L. cincinnatus; Lentinula edodes; Lentinus lepideus, L.giganteus, L. ponderosa, L. squarrosulus and L. tigrinus; Lentinulaspecies; Lenzites betulina; Lepiota rachodes and L. procera; Lepistanuda (=Clitocybe nuda); Lycoperdon lilacinum and L. perlatum; Lyophyllumdecastes; Macrocybe crassa; Marasmius oreades; Meripilus giganteus;Merulius incarnatus, M. incrassata and M. tremellosus; Morchellaangusticeps, M. crassipes and M. esculenta; Mycena citricolor, M.alcalina and M. chlorophos; Omphalotus olearius; Panellus stypticus, P.serotinus; Paxillus involutus; Phaeolus schweinitzii; Phellinusigniarius, P. pini, P. linteus and P. weirii; Pholiota nameko, P.squarrosa, Piloderma bicolor, Piptoporus betulinus; Pisolithustinctorius; Pleurotus citrinopileatus (=P. cornucopiae var.citrinopileatus), P. cystidiosus, (=P. abalonus, P. smithii), P. djamor(=P. flabellatus, P. salmoneostramineus), P. dryinus, P. eryngii, P.lignatilis, P. euosmus, P. nebrodensis, P. ostreatus, P. pulmonarius(=P. sajor-caju) and P. tuberregium; Pluteus cervinus; Polyporusindigenus, P. saporema, P. squamosus, P. tuberaster and P. umbellatus(=Grifola umbellata); Psathyrella hydrophila, Psilocybe allenii,aztecorum, P. azurescens, P. baeocystis, P. bohemica, P. caerulescens,P. coprophila, P. cubensis, P. cyanescens, P. hoogshagenii, P. mexicana,P. ovoideocystidiata, P. pelliculosa, P. semilanceata, P. serbica, P.subaeruginosa, P. tampanensis and P. weilii; Rhizopogon nigrescens, R.roseolus and R. tenuis (=Glomus tenuis); Schizophyllum commune;Schizopora paradoxa; Sclerocystis sinuosa; Serpula lacrymans and S.himantioides; Scleroderma albidum, S. aurantium and S. polyrhizum;Scutellospora calospora; Sparassis crispa and S. herbstii; Stereumcomplicatum and S. ostrea; Stropharia ambigua, S. aeruginosa, S. cyanea,S. albocyanea, S. caerulea, S. semiglobata, S. semigloboides, and S.rugosoannulata; Suillus cothurnatus; Talaromyces flavus; Termitomycesrobustus; Trametes elegans, Trametes T. gibbosa, T. villosa, T.cingulata, T. hirsuta, T. suaveolens and T. versicolor, Trichodermaviride, T. hamatum; Tricholoma giganteum and T. magnivelare (Matsutake);Tremella aurantia, T. fuciformis and T. mesenterica; Volvariellavolvacea; and numerous other beneficial fungi.

Preferred strains which have shown exceptional characteristics usefulfor the practice of this invention, include, by way of example but notof limitation, Fomes fomentarius (NY state), Ganoderma applanatum(Strain Duckabush), Fomitopsis officinalis (Strains I, VI, X),Fomitopsis pinicola (Strain I), Ganoderma oregonense (Meadow Lake),Heterobasidion annosum (Dosewallips), Pleurotus ostreatus (StrainsPW-OST, Nisqually), Psilocybe azurescens (Stamets strain), Strophariarugosoannulata (Strain F), Trametes versicolor (Kamilche Point) andInonotus obliquus (Stamets NY).

Additional suitable mushroom genera and species can be found in standardmycological field guides such as, but not limited to, MushroomsDemystified (1979, 1986) by David Arora, The Audubon Society Field Guideto North American Mushrooms (1981, 1995) by Gary Lincoff, and PsilocybinMushrooms of the World (1996) by Paul Stamets, Mushrooms of the PacificNorthwest (2009) by Steve Trudell and Joe Ammirati, and CaliforniaMushrooms: The Comprehensive Identification Guide by Dennis E.Desjardin, Michael G. Wood and Frederick A. Stevens. Continually updatedlists of suitable species based on the most recent DNA analysis can befound at the Tree of Life and Encyclopedia of Life (EOL) web projects.Other data bases include those maintained or referenced by theMycological Society of America, La Asociación Latinoamericana deMicologia, the European Mycological Association, the Asian MycologicalAssociation, and the International Mycological Association. Variousfungal DNA reference databases have been published by theseorganizations. One commonly used is MycoBank maintained by theInternational Mycological Association, which is useful for keeping upwith the many latest taxonomic changes and trends which are constantlyin flux as the science progresses.

The extracts from the mycelium of Fomitopsis officinalis particularly,and Inonotus obliquus, Fomitopsis pinicola, Fomes fomentarius, Ganodermaresinaceum and other species in the Polyporaceae generally, and extractsfrom gilled species such as Schizophyllum commune, reduce thepathogenicity of viruses to bees by directly reducing the viral particlepopulations while also fortifying the immune systems of bees, thuslimiting their virulence and transmissibility.

Moreover, bees better benefit from a combination of a mixture of theantiviral components generated by the mycelium with the antimicrobialproperties of coumarins and other compounds produced by the Fomitopsisofficinalis mycelium. The extracellular exudates secreted by themycelium of the beneficial fungi described herein have a combination ofthese constituents but balanced to have the net benefit of attractingbees, so they are fortified with immune enhancing, and nutritionallybeneficial constituents. This multifaceted effect results in fortifyingthe immune systems of bees and their colonies, making them lesssusceptible to viral, bacterial, protozoal, and fungal mitigateddiseases.

The present inventor has found that Ganoderma, Fomes, Fomitopsis,Fomitiporia, Ganoderma, Antrodia, Inonotus, Irpex, Lenzites, Phellinus,Sparassis, Hypholoma, Pleurotus, Schizophyllum, and Stropharia speciesdemonstrate strong anti-fungal properties and expects these will also beuseful for controlling fungal pathogens afflicting bees, including butnot limited to Nosema species and other pathogenic microsporidia,Chalkbrood and Stonebrood.

The aggressive wood rotting fungi listed in this application competewith many other fungi to establish their dominance in ecological niches.The polypore mushroom species, in particular species of Antrodia, Fomes,Fomitopsis, Ganoderma, Grifola, Heterobasidion, Inonotus, Stereum andTrametes, produce anti-fungal properties, present in extracts, whichthis inventor suggests will be effective against Nosema, amicrosporidium fungal parasite plaguing bees worldwide.

Moreover, the antibiotic effect of these extracts on microsporidium beeparasites, particularly Nosema apis, the cause of ‘Nosema,’ recentlyreclassified as a simple fungus, will prove to be a beneficialco-occurring factor.

Another advantage of the present invention is the wide-rangingantiviral, antibacterial and antifungal properties derived frommycelium. Many of the inventor's mycelium extract fractions demonstrateantiviral activity even when the bioguided fractionation pathway led toantibacterials.

Microbial agents are often thought of as microbial-type specific (thereis some cross-over between antibacterials and anti-parasitics and nowmay even be at least one class with both anti-bacterial and anti-fungalactivity), but considering how difficult it is to attain anti-viralspecificity alone, and the absence of known shared molecular targetsbetween bacteria and viruses that also exhibit any degree of selectivitywith respect to the host, broad anti-microbial activity is rare. Withoutbeing bound to any theory, the inventor would hypothesize that theextracts are acting as immunostimulators, immuno-potentiators andimmuno-regulators with antiviral, antibacterial and antifungal effects.

It is hypothesized that the mycelial components discussed above and/orother known and unknown compounds are anti-bacterial and anti-fungal,helping immunity, and hence the interaction between bees and extracts ofpure cultured mycelium within discrete concentrations is anunanticipated advantage of the present invention.

Hyphodermella corrugata, Polyporus umbellatus, and Piptoporus betulinusare species of the polyporales known to the author from his research toexhibit strong antiprotozoal properties. Agaric acid is thought to beone agent responsible for Piptoporus betulinus' anti-protozoal activity.Agaric acid is also produced by Fomitopsis officinalis, and possibly byother species in the polyporales. The production of acanthocytes byStropharia rugosoannulata, known to kill nematodes, may also provideantiprotozoal and anti-miticidal benefits to bees. As such, thesespecies and their relatives would be preferred for testing forantiprotozoal activity and up-regulation of antiprotozoal genes in bees.

Pesticides

As bees are limited in the number and variety of enzymes needed todenature natural and anthropogenic toxins, these toxins impair theirbaseline immunity, making them more susceptible to pathogens fromnumerous vectors—from Varrroa mites, Nosema and microsporidia fungi,Phorid flies, and the viruses and bacteria they carry. By increasing thebees' ability to degrade these toxins by up-regulation of morecytochrome P450 genes, GST genes and/or CCE genes, the bees' immunestate is improved to better resist these assaults and other stressfactors. Moreover, by providing bees with a blend of fungal extractsthat specifically limit the severity of assaults from Phorid flies,Varrroa mites, Nosema fungi and viruses, bee colony health can befortified for the long-term health of the brood, the workers, the queenand her drones. These fungal components are naturally incorporated intothe honey and propolis, thus imparting an advantage to developinggenerations. Ultimately, not only are bees are protected, but honeyproduction is expected to increase, and the quality of the honey bettersupports downstream generational health and survivability.

The inventor has isolated various strains of mushroom fungi, includingPleurotus ostreatus, Trametes versicolor, and Psilocybe azurescens thathave demonstrated superior abilities to “bioremediate” or“mycoremediate” various toxins including oil, pesticides and nerve gasessuch as Sarin, Soman and VX (dimethylmethylphosphonate), working withBattelle Laboratories, a public report of which was published in Jane'sDefence Weekly. Fungi could combat chemical weapons, Jane's DefenceWeekly, 32(7):37 1999. Those mushroom species useful in bioremediation(“mycoremediation”) of toxins, pollutants and pesticides and extracts oftheir mycelium are expected to contain various substances useful inturning on, up-regulating and modulating the genes necessary for thebiodegradation of pesticides. Since many such genes, or the systems suchas the cytochrome system, are evolutionarily conserved or similar, it isexpected that the extracts of the mycelium of such mushrooms willsimilarly be useful in up-regulating genes and systems in bees todegrade and deal with such pesticides. Useful and preferred speciesinclude the saprophytic mushrooms Pleurotus ostreatus and otherPleurotus species, Trametes versicolor, Trametes elegans and otherTrametes species, Fomes fomentarius, Fomitopsis officinalis and F.pinicola, Ganoderma lucidum, G. resinaceum, G. applanatum, G. annulare,G. brownii, G. collosum, G. lingzhi, G. curtisii, G. oregonense and G.tsugae; Heterobasidion annosum, Inonotus obliquus, I. hispidus, Irpexlacteus, Laetiporus sulphureus, L. conifericola, L. cincinnatus,Polyporus umbellatus, Polyporus elegans, Polyporus squamosus, Antrodiaspecies, Phaeolus schweinitzii, Boletus mirabilis, Gymnopus peronatus,Mycena alcalina, M. aurantiidisca, M. haematopus, Psilocybe azurescens,P. allenii, P. subaeruginosa, P. ovoideocystidiata, P. cubensis, P.cyanescens, Panaeolus cyanescens, Stropharia ambigua, Strophariarugosoannulata, Stropharia coronilla, Hypholoma capnoides, H.fasciculare, H. aurantiaca and other species in the Strophariodeae andStrophariaceae, Lenzites betulinus, Pholiota adiposa, Pholiotaterrestris, Pholiota nameko, Agrocybe aegerita, A. praecox, A. arvalis,Collybia tuberosa, Collybia, Psathyrella hydrophila, P. epimyces,Marasmius oreades, and their associated, numerous “satellite genera” aswell as the other gilled and polypore genera and species known to themycological science as primary and secondary decomposers of celluloseand lignin.

When not immunologically depressed from man-made and natural toxins,bees natural host defense can better protect bees from other deleteriousagents, including viruses and pathogens transmitted by Varrroa mites.

As our knowledge of the many derivatives of this overarching inventionexpands, the inventor anticipates that individual fungal species willoffer a unique set of benefits. Some will be more antiviral. Some willactivate the detoxification pathways in bees better than others againstdifferent toxins. Some emit fragrances greater in their attractiveproperties. As such, blends or “fungal cocktails” of species can becustomized according to the needs of the bees, the bee keepers, based ontheir desired targeted benefits, the ecosystem particulars, andconditioned upon the availability of basic materials.

For example, critical to the bee industry is the protection andgeneration of new queens. Queens are bred and reared by specialtybreeders who are at risk from mites transmitting the Black Queen CellVirus (BQCV). Finding a selective antiviral to protect queens is anothermajor advantage of this invention. For queen breeding and rearing, bothInonotus obliquus and Ganoderma resinaceum are very active antiviraladditives in reducing Black Queen Cell Virus (BQCV) but not as activeagainst Deformed Wing Virus (DWV), whereas other species are more activeagainst DWV. A blend of two or more mushroom species is thereforepreferred to provide a broad bioshield of antiviral activity to protectbees.

Levels of virions of the DWV may reduce the tensile strength of bees'wings that would limit their foraging range. Such weak wings would notbe visible to humans. Hive-mate bees often excommunicate DWV bees withnoticeable deformities. If the deformities were below the detectionlimits of ‘policing’ bees, these diseased bees would remain unobserved.At levels of this virus that would not be easily detected could resultin reducing tensile strength of bees whose wings would prematurely fail,especially during their foraging. This may partially explain why workerbees foraging times in the U.S. (and elsewhere) has been reduced toabout 4-5 days from historical averages of 9-10 days. Hence theinventor's extracts may improve overall wing strength of bee colonieswith great benefit to the beehive community.

Varroa Mites and Insect Parasites

While Varrroa mites are associated with the spread of viruses by actingas a viral reservoir and incubator, there is some indirect evidence thatreducing viral loads helps bees deal with Varroa infestation. Thearrival of Varrroa mites on Hawaii increased prevalence of DWV andmassively reduced DWV diversity, leading to the predominance of a singleDWV strain. Martin et al., Global Honey Bee Viral Landscape Altered by aParasitic Mite, Science, 336: 1304-1306, 8 Jun. 2012. This suggests somesort of co-evolutionary partnership; the virus may benefit Varroa mitesby causing problems in the developing bee pupae, by inhabiting andinfecting bee brains or by other mechanisms. Therefore, reducing DWVloads may both directly improve bee health and indirectly help bees dealwith Varroa. Bees with stronger immunity are better guardians of thehive. Healthy bees are commonly observed biting mites that are lockedonto other bees, to remove them or, even consuming infected larvae for‘recycling’ nutrients, leaving the Varroa without a host, making thefreed mite exposed to predation by the bees as it wanders in search of anew victim. Unfortunately for the bees, if the brood larvae have becomeinfected with viruses, this consumption behavior of worker bees exposesthem to the viruses already injected into the pupae. The bottom line isthat if viruses can be reduced while immunity is enhanced, thisinvention is a significant advancement for ensuring better bee colonyhealth.

The inventor has received several patents on compositions and methods ofusing the presporulating mycelium of entomopathogenic fungi as anattractant and treatment for controlling insects and arthropodsincluding mites, and the diseases insects and arthropods vector. Varroamites are known as a vector of the Israeli Acute Paralysis Virus and theTobacco Ringspot viruses. Varrroa mites, both plant and insect bitingmites, carry more than one virus or bacterial pathogen, meaning thatmites are one, albeit significant, vector carrying and introducingmultiple pathogens in the onslaught threatening beehive health. As beesweaken from viral exposure, for instance, they are less able to shed theattaching Varrroa mites. However, the mycelium and spores ofentomopathogenic fungi, particularly Aspergillus flavus, Metarhiziumanisopliae and Beauveria bassiana, can be used to attract, sicken orkill the Varrroa mites, reducing their activity, delivery of pathogenpayloads and numbers, thus tilting the balance in improving the hostdefense of the colony against CCD. Spores or mixtures of spores andmycelia of entomopathogenic fungi, including Metarhizium, Beauveria andthe Entomophthorales can similarly be used to sicken or kill Varrroamites, although mites may find spores repellant as compared topreconidial mycelium.

Moreover, extracts of Metarhizium anisopliae can be made specifically toattract, but not kill insects, including bees, by growing strains ofMetarhizium anisopliae that do not contain destructins, or have reducedlevels of these or other toxins, or reduced virulence and pathogenicity.Variability of toxins is true when comparing many strains of Aspergillusflavus, a known entomopathogenic fungus, primarily toxic due to itsaflatoxin content. Aflatoxin-free strains of Aspergillus flavus areavailable currently, which are naturally occurring or can be madethrough culture selections or genetic modifications. So too candestructin-free strains of Metarhizium anisopliae strains be created,selected for, or sourced from natural genomes. Strains can also beproduced which are not entirely free of destructins or aflatoxins, butproduce such low levels that they can be toxic to mites but not verytoxic to bees due to the fact that the bees' cytochrome P450 levels andpathways have been enhanced from exposure to coumaric acids and otherpolyphenols presented by the mycelium. In essence, the up-regulation ofcytochrome p450's (CYP's) may help bees better tolerate or detoxifydestructins or aflatoxins to which the bees are exposed from Metarhiziumanisopliae and Aspergillus flavus and other toxins produced byentomopathogenic fungi.

The advantage of a destructin-free or a reduced destructin strain ofMetarhizium anisopliae is that the extracts of the mycelium could beproduced with high sugar and terpene content, which would simultaneouslyattract bees and mites. Use of an appropriately sized mesh screen orbarrier or other means of selection allows for mites to be partitionedfrom bees so both bees and mites could be initially attracted to thesame location of the extracts (or similarly attracted to preconidialmycelium). The proportionality of the endemic entomopathogenic toxinscan be balanced to sicken mites but not bees. Using single or multiplefungal extracts as described herein offers a latitude and flexibility ofcustomized design, so that numerous devices, delivery systems,compositions and methods can be made available for the first time tofavor bee health and decrease CCD. Phorid flies, gnats and mitespredating on mushrooms are well known to the mushroom industry. What wasnot known is that extracts of entomopathogenic fungi prior tosporulation are attractive to these insects and arthropods. The presentinventor does not believe that hydroethanolic extracts of mushrooms ormushroom mycelium with these attractive properties were known to themushroom industry prior to this inventor's disclosures in pending andapproved patents.

Combining extracts of mushroom mycelia and oxalic acid with sugarenriched water loaded with spores or preconidial mycelia ofentomopathogenic fungi such as Metarhizium anisopliae and Beauveriabassiana will improve the miticidal actions of the combination of oxalicacids and entomopathogenic fungi and the anti-miticidal properties ofother components resident or added to sugar water, pollen patties or beesprays, for instance.

However, oxalic acid is reactive to the minerals in the fungal extracts,and this may possibly pose a hurdle for effective formulation. Whencombining oxalic acids with the extracts of filamentous Basidiomycetesfungi, the resident minerals (calcium, phosphorus, iron) may possiblybind with the oxalic acid thus reducing the mineral scouring, miticidalpotential of the oxalic acids. Therefore, if such is a problem,demineralization of the fungal extracts before combining oxalic acid tothe fungal extracts is an embodiment of this invention. Demineralizationemploys any of numerous methods useful for demineralizing of the fungalextracts so as to prevent conversion of the reactive oxalic acid intowater insoluble salts by eliminating calcium and other minerals residentwithin the fungal extracts. One method of many available is to make useof ion exchange resin technologies. The fungal extracts can be added todistilled water at a ratio of 1:10 preferably, with ranges of 1:1 beingthe most concentrated and 1:100 being most dilute but less preferable.Upon completion, minerals in the fungal extracts, which might otherwiseneutralize the anti-miticidal properties of oxalic acid, will be largelyif not completely removed. Thereupon, oxalic acid can be added to thereduced mineral, fungal extracts in a sufficient quantity to have ananti-miticidal effect, in the range of 1-10% of oxalic acid to the massof the solution, resulting in a low pH in the 0.5-3.5 pH range, with anoptimal range in the 0.5-2.0 pH range.

Ganoderma lucidum is one of the species of particular interest (alongwith Ganoderma resinaceum, Ganoderma applanatum, Ganoderma brownii,Ganoderma curtisii, Ganoderma oregonense, Ganoderma tsugae, Ganodermalingzhi, Ganoderma capense, Ganoderma annularis, and Ganoderma collosum)to the inventor as it not only has strong antiviral properties, but hascomplexes of sugars that result in its mycelium producing a viscoussyrup-like “mycological honey” that can be used to help bees surviveCCD. The inventor and his team at Fungi Perfecti, LLC have also notedthat the extracts of Ganoderma resinaceum will not freeze, even whenfreeze driers achieve temperatures less than −50° C. under high vacuum,whereas species tested outside the genus Ganoderma readily freeze driedinto a dried state under the same conditions. The inventor hypothesizesthe mycelial extract of Ganoderma resinaceum, and likely extracts ofrelated Ganoderma species, maintains a liquid state even under cryogenicconditions due to its unique assortment of complex sugars, sterols, andglycoproteins binding to form a unique liquid matrix far different thanany other species tested. This extract may have the potential to improvebee and colony winter survival and potential as an anti-freeze withbroad reaching implications for medicine, avionics, space travel, andusefulness under extreme temperature conditions for lubricating,preservation, and extremophile chemistry.

Since extracts of mycelium of Ganoderma resinaceum and Inonotus obliquusgrown on sterilized rice reduce the Black Queen Cell Virus by >500:1,and extracts of mycelium of Ganoderma resinaceum grown on sawdust reducethe Lake Sinai virus by >500:1, and extracts of mycelium of Fomesfomentarius grown on sawdust reduce the Deformed Wing Virus by >1000:1,combinations of these extracts that are grown on sterilized substratescan be presented to bees, utilizing a 1:1:1 or other ratio, in theirfeed or sugar water to create a broad armamentarium of antiviraldefenses to help bees fend off a plethora of pathogenic viruses andother pathogens, including but not limited to mites, other insects,bacteria, fungi, thereby extending lifespan, foraging ranges, stamina,pollination services, and their overall ability to withstand many of thestressors which are part of the cause of colony collapse disorder. Dronebees not infected with viruses are less likely to wander into native beecolonies, limiting cross infection between bee species. Moreover, theuse of a multiplicity of these aforementioned extracts can help activatedetoxification pathways allowing the bees to better survive toxinexposures, improve their microbiomes and hive behavior. Many beneficialgene expressions from bees can result as a direct consequence of thisinvention.

In essence, these extracts create a symphony of benefits that help theoverall immunity of bees, their hives, their progeny, and the manybenefits to plant communities, especially those used in agriculture.Given the species-specificity factors of each fungal species used,combinations are not limited to those that are disclosed herein but manyother fungal species are anticipated to be useful. Hence, uniquecombinations can be devised to create specific formulas. Thesediscoveries can be ‘dialed’ in for formula optimization considering thethreats bees encounter. By combining these extracts with other beeremedies, this invention can enable those remedies to work moreeffectively.

In all of the following examples, the inventor anticipates, asderivatives of his discovery, that bioguided fractionation methods willlead to increasing the potency, increasing efficacy, and reducing thecost of production, manufacturing, and the implementation of saidinventions and its many elaborations, which become obvious subsequent tothis paradigm shifting discovery.

EXAMPLES Example 1

Fomes fomentarius, Fomitopsis officinalis, Fomitopsis pinicola,Ganoderma resinaceum, Inonotus obliquus, Piptoporus betulinus, Trametesversicolor, Schizophyllum commune and other mushroom species arecultured utilizing any known means for inoculating and means forcultivating medicinal mushroom mycelium or means for growing mycelium onrice, barley, flaxseeds or other grains, agricultural debris, or forestproducts such as sawdust or wood chips (for a list of substrates and adiscussion of inoculating and cultivating mushroom mycelium, SeeStamets, Growing Gourmet and Medicinal Mushrooms, 1993, Ten Speed Press,Berkeley, Calif., and Stamets & Chilton, The Mushroom Cultivator,Agarikon Press, Olympia, Wash.). Liquid inoculation is preferred forgrain substrates, although inoculation with colonized agar may beutilized, and inoculation with colonized grain is preferred for sawdustor wood chip substrates. When the mycelium reaches a dense mass ofgrowth (preferably after 20 but before 120 days growth in fermentationor in solid state fermentation subsequent to inoculation, but wellbefore fruitbody formation) mycelial mass can be extracted throughsimple aqueous, water/ethanol (both of which are preferred) or ethanolwashing of the substrate, or from compression of the substrate, or othermeans for extracting discussed herein, all of which will result in aliquid fluid or capture-able extract including extracellular exudates.These extracts can be utilized as they are, or alcohol (25-50% byvolume) may be added to aqueous extracts as both a preservative andsolvent (which will precipitate water-soluble polysaccharides). Thehydroethanolic extract can be evaporated or removed, or the alcohol andwater may be evaporated and removed separately. The crude extract can becell free filtered using a 0.12-0.20 μm filter. This extract can befrozen or dried for future use. Alternatively, non-aqueous ornon-ethanolic solvent extracts such DMSO, ethyl acetate, ether, or“edible” solvents such as 3-methoxy-3-methyl-1-Butanol (MMB), PEG-400,glycerol and propylene carbonate or other alcohols or solvents orcombinations of solvents known to the art may be utilized, orsubcritical or supercritical fluid extracts utilizing, for example,carbon dioxide or water, and optional co-solvents such as alcohols, maybe utilized, or microwave-assisted extracts may be utilized. Extractsmay also be prepared via steam distillation of volatile components,similar to the preparation of “essential oils” from flowers and herbs.Suitable alcohols include those containing from 1 to 10 carbon atoms,such as, for example, methanol, ethanol, isopropanol, n-propanol,n-butanol, 2-butanol, 2-methyl-1-propanol (t-butanol), ethylene glycol,glycerol, etc. Suitable organic solvents include unsubstituted organicsolvents containing from 1 to 16 carbon atoms such as alkanes containingfrom 1 to 16 carbon atoms, alkenes containing from 2 to 16 carbon atoms,alkynes containing from 2 to 16 carbon atoms and aromatic compoundscontaining from 5 to 14 carbon atoms, for example, benzene, cyclohexane,cyclopentane, methylcyclohexane, pentanes, hexanes, heptanes,2,2,4-trimethylpentane, toluene, xylenes, etc., ketones containing from3 to 13 carbon atoms such as, for example, acetone, 2-butanone,3-pentanone, 4-methyl-2-pentanone, etc., ethers containing from 2 to 15carbon atoms such as t-butyl methyl ether, 1,4-dioxane, diethyl ether,tetrahydrofuran, etc., esters containing from 2 to 18 carbon atoms suchas, for example, methyl formate, ethyl acetate, butyl acetate, etc.,nitriles containing from 2 to 12 carbon atoms such as, for exampleacetonitrile, proprionitrile, benzonitrile, etc., amides containing from1 to 15 carbon atoms such as, for example, formamide,N,N-dimethylformamide, N,N-dimethylacetamide, etc., amines andnitrogen-containing heterocycles containing from 1 to 10 carbon atomssuch as pyrrolidine, 1-methyl-2-pyrrolidinone, pyridine, etc., halogensubstituted organic solvents containing from 1 to 14 carbon atoms suchas, for example, bromotrichloromethane, carbon tetrachloride,chlorobenzene, chloroform, 1,2-dichloroethane, dichloromethane,1-chlorobutane, trichloroethylene, tetrachloroethylene,1,2-dichlorobenzene, 1,2,4-trichlorobenzene,1,1,2-trichlorotrifluoroethane, etc., alkoxy, aryloxy, cycloalkyl, aryl,alkaryl and aralkyl substituted organic solvents containing from 3 to 13carbon atoms such as, for example, 2-butoxyethanol, 2-ethoxyethanol,ethylene glycol dimethyl ether, 2-methoxyethanol, 2-methoxyethyl ether,2-ethoxyethyl ether, etc., acids containing from 1 to 10 carbon atomssuch as formic acid, acetic acid, trifluoracetic acid, etc., carbondisulfide, dimethyl sulfoxide (DMSO), nitromethane and combinationsthereof. Extracts may also be prepared via sequential extraction withany combination of the above solvents or methods mentioned herein. Theextracts may be further refined by means known to the art to a morepotent antiviral form or an active pharmaceutical ingredient.

The extract can be added to any form of feed stocks for bee consumptionutilizing known means for adding and mixing liquids or liquids andsolids. The original extract can be used directly or diluted and addedto drinking water, sugar water, bee candy, honey, propolis, pollenpatty, grease patty and protein supplements to give improved bee feedsand nutritional products and improved pollen supplements, dietarysupplements, feeding supplements and nutritional supplements. Theextracts may also be incorporated into sprays and used with means forspraying to produce “extract treated” bees, beehives and beehivecomponents including frames, supers and wax foundations or used withmeans for spraying and treating areas to be pollinated by bees, the areasurrounding beehives or areas frequented by wild bees. The supernatantextracts may be added to the sugar water or other feed water, to beepatties, bee bread, propolis, or in any way to enable bees to makecontact with these longevity extending mycelial extracts. Moreover, theextracts can be added to means for killing mites such as oxalic acid orother miticides for topical application and ease of use. Ingestion andcontact by bees improve the bees' ability to build immunity throughup-regulating of toxin degrading enzymes, reduces pathogen payloads andprovide a healthy source of diverse sugars, amino acids, vitamin B's,and nutrients. Moreover, the precipitate, although partitioned from thesupernatant, contains within it nutrient rich, and antiviral, healthsupporting properties, which can be used also as feedstock forbenefitting bees. Both the supernatant and the precipitate can becombined, and enzymatically converted using amylase and other enzymes tofurther transform starches and other ingredients into a more effectivecomposition.

Exemplary compositions comprise one or more fungal extracts in anaqueous or ethanol solvent and one or more sugars, carbohydrate sources,flavors, colorants, sweeteners, thickeners, or preservatives, whereinthe composition is a tincture, elixir, or dried, solvent free dosageform. The extracts of the present invention may also optionally beenhanced by use of protectants and nutrients (sugars or carbohydratesare preferred materials that have both protectant and nutrientqualities), and materials such as wetting agents, surfactants andsurface active agents, dispersants, emulsifiers, tackifiers oradhesives, penetrants, fillers, carriers, antibiotics or nutritionalsupplements, dispersants, emulsifiers, humectants, arrestants, feedingstimulants, sex pheromones, aggregating pheromones, trail pheromones,encapsulating materials and combinations thereof.

Additional pharmaceutical excipients useful for the compositions asdescribed herein include, for example, the following: Acidifying agents(acetic acid, glacial acetic acid, citric acid, fumaric acid,hydrochloric acid, diluted hydrochloric acid, malic acid, nitric acid,phosphoric acid, diluted phosphoric acid, sulfuric acid, tartaric acid);Alkalizing agents (ammonia solution, ammonium carbonate, diethanolamine,diisopropanolamine, potassium hydroxide, sodium bicarbonate, sodiumborate, sodium carbonate, sodium hydroxide, trolamine); Antifoamingagents (dimethicone, simethicone); Antimicrobial preservatives(benzalkonium chloride, benzalkonium chloride solution, benzethoniumchloride, benzoic acid, benzyl alcohol, butylparaben, cetylpyridiniumchloride, chlorobutanol, chlorocresol, cresol, dehydroacetic acid,ethylparaben, methylparaben, methylparaben sodium, phenol, phenylethylalcohol, phenylmercuric acetate, phenylmercuric nitrate, potassiumbenzoate, potassium sorbate, propylparaben, propylparaben sodium, sodiumbenzoate, sodium dehydroacetate, sodium propionate, sorbic acid,thimerosal, thymol); Antioxidants (ascorbic acid, ascorbyl palmitate,butylated hydroxyanisole, butylated hydroxytoluene, hypophosphorousacid, monothioglycerol, propyl gallate, sodium formaldehyde sulfoxylate,sodium metabisulfite, sodium thiosulfate, sulfur dioxide, tocopherol,tocopherols excipient); Buffering agents (acetic acid, ammoniumcarbonate, ammonium phosphate, boric acid, citric acid, lactic acid,phosphoric acid, potassium citrate, potassium metaphosphate, potassiumphosphate monobasic, sodium acetate, sodium citrate, sodium lactatesolution, dibasic sodium phosphate, monobasic sodium phosphate);Chelating agents (edetate disodium, ethylenediaminetetraacetic acid andsalts, edetic acid); Coating agents (sodium carboxymethylcellulose,cellulose acetate, cellulose acetate phthalate, ethylcellulose, gelatin,pharmaceutical glaze, hydroxypropyl cellulose, hydroxypropylmethylcellulose, hydroxypropyl methylcellulose phthalate, methacrylicacid copolymer, methylcellulose, polyvinyl acetate phthalate, shellac,sucrose, titanium dioxide, carnauba wax, microcrystalline wax, zein);Colorants (caramel, red, yellow, black or blends, ferric oxide);Complexing agents (ethylenediaminetetraacetic acid and salts (EDTA),edetic acid, gentisic acid ethanolamide, oxyquinoline sulfate);Desiccants (calcium chloride, calcium sulfate, silicon dioxide);Emulsifying and/or solubilizing agents (acacia, cholesterol,diethanolamine (adjunct), glyceryl monostearate, lanolin alcohols, mono-and di-glycerides, monoethanolamine (adjunct), lecithin, oleic acid(adjunct), oleyl alcohol (stabilizer), poloxamer, polyoxyethylene 50stearate, polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil,polyoxyl 10 oleyl ether, polyoxyl 20 cetostearyl ether, polyoxyl 40stearate, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate80, diacetate, monostearate, sodium lauryl sulfate, sodium stearate,sorbitan monolaurate, sorbitan monooleate, sorbitan monopalmitate,sorbitan monostearate, stearic acid, trolamine, emulsifying wax);Filtering aids (powdered cellulose, purified siliceous earth); Flavorsand perfumes (anethole, benzaldehyde, ethyl vanillin, menthol, methylsalicylate, monosodium glutamate, orange flower oil, peppermint,peppermint oil, peppermint spirit, rose oil, stronger rose water,thymol, tolu balsam tincture, vanilla, vanilla tincture, vanillin);Humectants (glycerol, hexylene glycol, sorbitol); Plasticizers (e.g.,castor oil, diacetylated monoglycerides, diethyl phthalate, glycerol,mono- and di-acetylated monoglycerides, propylene glycol, triacetin,triethyl citrate); Polymers (e.g., cellulose acetate, alkyl celluloses,hydroxyalkyl, acrylic polymers and copolymers); Solvents (acetone,alcohol, diluted alcohol, amylene hydrate, benzyl benzoate, butylalcohol, carbon tetrachloride, chloroform, corn oil, cottonseed oil,ethyl acetate, glycerol, hexylene glycol, isopropyl alcohol, methylalcohol, methylene chloride, methyl isobutyl ketone, mineral oil, peanutoil, propylene carbonate, sesame oil, water for injection, sterile waterfor injection, sterile water for irrigation, purified water); Sorbents(powdered cellulose, charcoal, purified siliceous earth); Carbon dioxidesorbents (barium hydroxide lime, soda lime); Stiffening agents(hydrogenated castor oil, cetostearyl alcohol, cetyl alcohol, cetylesters wax, hard fat, paraffin, polyethylene excipient, stearyl alcohol,emulsifying wax, white wax, yellow wax); Suspending and/orviscosity-increasing agents (acacia, agar, alginic acid, aluminummonostearate, bentonite, purified bentonite, magma bentonite, carbomer,carboxymethylcellulose calcium, carboxymethylcellulose sodium,carboxymethylcellulose sodium 12, carrageenan, microcrystalline andcarboxymethylcellulose sodium cellulose, dextrin, gelatin, guar gum,hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethylcellulose, magnesium aluminum silicate, methylcellulose, pectin,polyethylene oxide, polyvinyl alcohol, povidone, alginate, silicondioxide, colloidal silicon dioxide, sodium alginate, tragacanth, xanthangum); Sweetening agents (aspartame, dextrates, dextrose, excipientdextrose, fructose, mannitol, saccharin, calcium saccharin, sodiumsaccharin, sorbitol, solution sorbitol, sucrose, compressible sugar,confectioner's sugar, syrup); Surfactants (simethicone); Tablet binders(acacia, alginic acid, sodium carboxymethylcellulose, microcrystallinecellulose, dextrin, ethylcellulose, gelatin, liquid glucose, guar gum,hydroxypropyl methylcellulose, methylcellulose, polyethylene oxide,povidone, pregelatinized starch, syrup); Tablet and/or capsule diluents(calcium carbonate, dibasic calcium phosphate, tribasic calciumphosphate, calcium sulfate, microcrystalline cellulose, powderedcellulose, dextrates, dextrin, dextrose excipient, fructose, kaolin,lactose, mannitol, sorbitol, starch, pregelatinized starch, sucrose,compressible sugar, confectioner's sugar); Tablet disintegrants (alginicacid, microcrystalline cellulose, croscarmellose sodium, crospovidone,polacrilin potassium, sodium starch glycolate, starch, pregelatinizedstarch); Tablet and/or capsule lubricants (calcium stearate, glycerylbehenate, magnesium stearate, light mineral oil, sodium stearylfumarate, stearic acid, purified stearic acid, talc, hydrogenatedvegetable oil, zinc stearate); Thickening agents (gelatin having a Bloomstrength of 50-100); Tonicity agent (dextrose, glycerol, mannitol,potassium chloride, sodium chloride); Vehicle: flavored and/or sweetened(aromatic elixir, compound benzaldehyde elixir, iso-alcoholic elixir,peppermint water, sorbitol solution, syrup, tolu balsam syrup); Vehicle:oleaginous (almond oil, corn oil, cottonseed oil, ethyl oleate,isopropyl myristate, isopropyl palmitate, mineral oil, light mineraloil, myristyl alcohol, octyl dodecanol, olive oil, peanut oil, persicoil, sesame oil, soybean oil, squalane); Vehicle: solid carrier (sugarspheres); Vehicle: sterile (Bacteriostatic water for injection,bacteriostatic sodium chloride injection); Viscosity-increasing (seesuspending agent); Water repelling agent (cyclomethicone, dimethicone,simethicone); and/or solubilizing agent (benzalkonium chloride,benzethonium chloride, cetylpyridinium chloride, docusate sodium,nonoxynol 9, nonoxynol 10, octoxynol 9, poloxamer, polyoxyl 35 castoroil, polyoxyl 40, hydrogenated castor oil, polyoxyl 50 stearate,polyoxyl 10 oleyl ether, polyoxyl 20, cetostearyl ether, polyoxyl 40stearate, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate80, sodium lauryl sulfate, sorbitan monolaurate, sorbitan monooleate,sorbitan monopalmitate, sorbitan monostearate, tyloxapol). This list isnot meant to be exclusive, but instead merely representative of theclasses of excipients and the particular excipients that may be used inoral dosage forms as described herein. Any methods known to the art andmeans for formulating extracts or active principal ingredients intoliquid or solid bee nutrients or feeding supplements or sprays or meansfor treating bees, means for drenching bees, means for dousing bees,means for “bathing” bees or means for wetting bees may be utilized.

The process steps of inoculating and cultivating a mushroom mycelium ongrain or sawdust and extracting the mycelium and mixing the extract withwater, sugar and optional ingredients are preferred. The process ofextracting with aqueous ethanol or the process of extracting with waterand adding ethanol is preferred.

Example 2

The medicinal mushroom mycelium is grown utilizing liquid culturetechniques. Whereas growing on rice might have 30-40% conversion of riceto mycelium, liquid vat culture may have essentially complete conversionwith >3× more mycelium per unit mass. Hence the liquid vat culture ofmycelium and its extracellular metabolites will be easier to utilize inthe development of this invention as the process of using vat cultureeliminates the need to remove non-metabolized substrate ingredients. Anymeans for inoculating and cultivating mushroom mycelia via liquidculture may be utilized.

Example 3

Mycelial extracts can be made in many ways. One preferred method formaking the mycelial extract is to grow mycelium on a mixture containingequal volumes of sterilized or fermented grain (barley, flaxseed, rice,oats, millet, wheat, rye, corn, seeds, including nuts, sawdust or woodchips (Douglas fir, pines, oaks, birches, alders, aspens, cottonwoods,olives, Prunus trees)), that has been incubated for more than 4 weeksbut less than 20 weeks, and then immerse this myceliated mass into a anequal volume of a 50:50 water-ethanol solution, adding the ethanolfirst. Allow to sit at room temperature for two weeks, and then drainthe liquid and press to expel the liquid extract contained within themycomass. Over several days, a precipitate will fall out of thehydroethanolic solution. The hydroethanolic supernatant is drawn offabove the pasty precipitate. After several more weeks, or by using acentrifuge, the precipitate further concentrates into a semisolid state.The supernatant is preferably utilized with means for feeding bees ormeans for spraying or treating bees to improve bee longevity and reduceantiviral loads. However, the precipitant also contains compounds usefulfor improving immunological health.

These precipitated wet semisolids are removed and heated to 50° C. for6-8 hours while stirring. The wet volume of semisolids is reduced toabout 40% of the original wet semisolids. The drying down of thesemi-solids into the caramel “honey-like” substance yields about 16% ofthe original wet solids wet. Therefore, using 1000 mL of wet solids(which was 40% of the initial extract) yields about 170 mL (within arange of 100-200 mL) of thick syrupy caramel like substance. Continuedheating and stirring concentrates this substance with noticeably sweeterproperties. Amylase, chitinase or other enzymes may be optionally added.The extract can be crystallized, powdered, and used as amendment toother treatments. The liquid, semisolid and crystallized forms arenoticeably sweet in taste and could be considered a medicinal candy-likesubstance useful to both bees and people in a wide number ofapplications.

Example 4

A mycelial extract is made utilizing means for extracting fruitbodies ormycelium of basidiomycetous fungi including Ganoderma resinaceum in hotwater (80-100° C.) for several hours and combined with the roomtemperature (10-30° C.) water extraction of Fomes fomentarius,Fomitopsis officinalis, Fomitopsis pinicola, Ganoderma resinaceum,Inonotus obliquus, Piptoporus betulinus, Trametes versicolor and/orSchizophyllum commune mycelium grown on grain or wood. To these waterextracts, ethanol is added to make the solution greater than 22% EtOH(ethanol), preferably 35-45% EtOH, precipitating polysaccharides out ofsolution, which settle at the bottom of the extraction vessel. Upondrawing off the supernatant, the precipitated polysaccharides, rich inglycosides, glycoproteins and other ‘nectar-like’ nutrients, arecollected and heated between 50-70° C. over several hours, resulting inthe creation of a sweet residue attractive to and beneficial to bees.Optionally enzymes such as amylase and chitinase can be incorporated toenhance the sweetness, attractiveness, palatability, and medicinal valueof this mixture. Alternately, the supernatant can be stored over severaldays, which further yields useful precipitating polysaccharides. Theseprecipitates contain complex sugars, antivirals, antibacterials,cytochrome p450 up-regulating coumaric acids and coumarins, and can becombined with other ingredients used in the feeding water, pollenpatties, propolis, bees wax, sprays, or in any delivery system wherebybees make contact with these precipitates, helping bees overcomestressors associated with colony collapse disorder.

Example 5

A mycelial extract made from extracting fruitbodies or mycelium, ormyceliated grain or sawdust of basidiomycetous fungi including Ganodermaresinaceum is first soaked in 100% ethanol (1:1 ratio by mass) for 1-7days. Upon draining off the ethanol, the mushroom- or mycelial-marc isimmersed into hot water (80-100° C.) for several hours and combined withthe room temperature (10-30° C.) water immersion and extraction of Fomesfomentarius, Fomitopsis officinalis, Fomitopsis pinicola, Ganodermaresinaceum, Inonotus obliquus, Piptoporus betulinus, Trametes versicolorand/or Schizophyllum commune mycelium grown on grain or wood. To thesewater extracts, the ethanol extracts previously described are added tomake the total combined solution greater than 22% EtOH, preferably35-45% EtOH. Upon addition of ethanol fraction, polysaccharidesprecipitate out of solution and settle at the bottom of the extractionvessel. The supernatant is preferred for antiviral activity and lifeextension in bees.

The precipitate also holds p-coumaric acids, and additionally othernutrients, which can be used to feed bees. These p-coumaric enrichedprecipitates also contain complex sugars, antivirals, antibacterials,and families of coumarins, and can be combined with other ingredients,such as the water soluble mushroom polysaccharides, corn syrup or sugarsused in sweetening the feeding water, or additionally incorporated as aningredient in pollen patties, propolis, bees wax, sprays, or in anydelivery system whereby bees make contact with these precipitates,helping bees overcome stressors associated with colony collapsedisorder.

p-coumaric acid, being more soluble in ethanol than water, is richer inthe ethanolic extracted supernatant. (The ethanolic supernatant, withconcentrated p-coumaric acids, is a reservoir of bee-beneficial p450coding compounds.) This hydroethanolic supernatant can be stored overseveral days, which further yields a mixture of polysaccharides, butwhich is proportionately higher in p-coumaric acids than the hot waterfractions alone.

Example 6

Once the extracellular metabolites in the supernatant from pure culturedmycelium of Fomes, Fomitopsis, Ganoderma, Inonotus, Trametes,Schizophyllum species, or other antiviral, bee-benefitting fungi, areextracted from the pure cultured mycelial scaffolding, these mycelial‘juices’ can be preserved in alcohol to prevent immediate souring. Theseantiviral liquids can be put in freezers, subzero freezers, oralternatively, using a rotary evaporator, the hydroethanolic extractscan be reduced to a near solid or solid state, in essence a paste, andsealed inside of a sterile container for ease of transport and use. Thispaste can be re-solubilized directly back into the bee sugar feed water,or back into alcohol to decrease viscosity, according to the preferencesof the end-user beekeeper. The process steps of extracting the mycelium,preserving with ethanol or other preservative and, optionally, removingthe solvent are preferred.

Example 7

The mycelia of antiviral-active fungal species can be cultivated on arange of mixtures of grain and sawdust. A preferred range would be a 50%sawdust and 50% mixture grain or grain spawn to a range of 90% sawdustand 10% grain or grain spawn, both of which is balanced to have a 35-55%moisture content to create dense “bricks” of mycelium. (Alternatively,sawdust can be replaced with cereal straw or other plants-basedingredients can be utilized.) After incubating for at least 7 days,preferably 30-160 days, these mycelial bricks can be soaked in water,and the water extracted compounds immediately put into an equal mass ofalcohol, rendering the mixture to have more than 30% ETOH. Above ˜22%ETOH, beta glucans and other polysaccharides precipitate out. The“marc”—the original substrate material used prior to soaking withwater—may be reused for repetitive extracts. Moreover, theseprecipitants as well as the marc, with the addition of amylases andchitinases, can be converted into a nutritious honey like syrup or pasteuseful as a nutraceutical or as a functional food for enhancing immunityof animals, including but not limited to bees, birds and humans. Thisformulation for making antivirals is unique as mycelium produced forspawn production is typically added to a sawdust substrate between 2% to<10% of total mass and the incubating time is classically 5-14 days. SeeGrowing Gourmet & Medicinal Mushrooms, 1993, Ten Speed Press. Five tofourteen days is the preferred duration of incubation of mycelium asstated by the largest spawn producer in the world, Sylvan Inc. Spawnpast this date is unusable commercially and is discarded as standardpractice. Amateurs may use older spawn but much beyond this range isdifficult to use as the myceliated grain kernels or sawdust particlesglom together during prolonged incubation of mycelium. (A measure ofgood spawn quality is the ability of it to separate into individualparticles (kernels or fragments), maximizing multiple points ofinoculation and re-growth potential.) Older spawn when shaken, bruises,clumps together and the resulting necrotic tissue is highly susceptibleto contamination. Moreover, the spawn loses vitality and colonizationonto new substrates is diminished in comparison. The process used herefor the antiviral and longevity enhancing extracts uses aged myceliumimmediately immersed the into a hydroethanolic solution before regrowthand co-occurring contamination result. Hence this process is unobviousto the mindset of conventional spawn producers. In contrast, theincubating mycelium for making extracts described in this inventionextends well past the window of usefulness as spawn, being incubated onsawdust for >45 but <120 days, and on grain for >20 but <60 days withinwhich is a preferred time period for making extracts from the myceliatedsubstrate (the strain, species, spawning rates, substrate mass, depth,incubation temperature, periodic shaking, gas exchange and light allinfluence myceliation and can be adjusted to increase extract potency).All factors being equal, one preferred method is to increase thespawning rate greater than conventional practices utilizing >10%inoculation rate onto sawdust or grain with pure cultured spawn. Underprolonged incubation of a few months, the grain more quickly decomposes,leaping off into the sawdust and as result unusually dense colonizationoccurs throughout the grain/sawdust substrate. The quick decompositionreleases a cascade of enzymes, which encourages subsequent regrowth.This moist mycomass can then be compressed and the extracellularexudates and intracellular metabolites collected in liquid form. Storagein the dark or with controlled exposure to blue light in the 300-400nanometer wavelength can enhance antiviral, detoxification and longevityenhancing benefits.

Additionally, the substrate mass can be disturbed in between eachsoaking to enhance extracellular metabolite production. As an example:the remaining now compressed mycomass can then be immersed in an equalmass of water for 10-60 minutes, which will result in it swelling withwater which re-initiates the growth process. Subsequent to eachcompression or extraction, after the water is removed, the grain/sawdustmass is re-transferred into incubation bags, incubated for severalweeks, and soaked again one week to four weeks later, depending uponspecies, strains, and other growth factors. This ‘milking of themycelium’ greatly extends and improves yield production of antiviralextracts. This process can be repeated multiple times. Maltose, sucrose,glucose, fructose, xylose, arabinose, galactose, rhamnose other sugarsand essential nutrients (peptone, soy protein, yeasts, bacteria,minerals) can be added to the water used for soaking to enhanceregenerative growth of the mycelium and its suites of extracellular andintracellular metabolites. Such regeneration methods also allow forimmunizing the mycelium, epigenetically, by exposing it to low titers ofspecific viruses or other pathogens, activating antimicrobial genesequences, and thereby increasing anti-pathogen properties, speciesspecifically.

Example 8

For each type of aqueous ethanolic supernatant mushroom myceliumextract, prepared from mycelium grown on grain and extracted accordingto Example 3 at room temperature, mixed aged honey bees from a singlehive were collected on a single day and distributed at random into 16cages of roughly 100 bees each. Each set of 16 consisted of four controlcages (fed sugar syrup), four low concentration cages (fed myceliumextract in sugar syrup at 0.1% v/v), four medium concentration cages(fed mycelium extract in sugar syrup at 1% v/v), and four highconcentration cages (fed mycelium extract in sugar syrup at 10% v/v). Ineach group of four cages, three cages were used for longevity tests andthe remaining replicate cage was used for total viral particle testing.A separate experiment was conducted to evaluate the effect of fungalextracts on specific virus types.

Improvement in Longevity

For longevity (survivorship) testing, each replicate (three cages foreach feeding concentration and control group) was monitored daily anddead bees were counted. For every day of the experiment, the totalnumber of bees that died as of that date was tabulated for eachreplicate cage for each fungal extract and for the control groups. Thesedaily dead bee tabulations were then used to calculate the percent ofthe original bees that were still surviving at each day of theexperiment. The mean percent survival rates were then calculated basedon the data from the three replicate cages for each fungal extract andfor the control group.

Survival plots were generated with time measured in days as theindependent variable (x-axis), and the mean percent of bees surviving atany point in the experiment as the dependent variable (y-axis); thelongevity graphs represent the % of the original population that issurviving at various points in time. See FIG. 1-4 . FIG. 1 is a linegraph showing percent survival of bees over time in days when givenextracts of the mycelium of Inonotus obliquus (0.1%, 1%, and 10% asrespectively shown by dotted, dashed and double-dash lines) with sugarwater as compared to a control population fed sugar water only (shown bya solid line). FIG. 2 is a line graph showing the percent survival ofbees over time in days when given extracts of the mycelium of Ganodermaresinaceum (0.1%, 1%, and 10% as respectively shown by dotted, dashedand double-dash lines) with sugar water as compared to a controlpopulation fed sugar water only (shown by a solid line). FIG. 3 is aline graph showing the percent survival of bees over time in days whengiven extracts of the mycelium of Fomitopsis pinicola (0.1%, 1%, and 10%as respectively shown by dotted, dashed and double-dash lines) withsugar water as compared to a control population fed sugar water only(shown by a solid line). FIG. 4 is a line graph showing the percentsurvival of bees over time in days when given extracts of the myceliumof Fomes fomentarius (1% shown by a light solid line) with sugar wateras compared to a control population fed sugar water only (shown by adark solid line). These and similar approaches can be used in thepractice of the invention to demonstrate the effect of the invention onthe health and longevity of the bees. The figures are mean values of theexperiments; the standard deviation bars were removed for clarity.

Some, but not all, of the results in these preliminary experiments werestatistically significant; improved results are expected in continuingtrials with more replicates. Statistical significance was assessed usingKaplan-Meier (product-limit) survival estimates prepared using JMP®statistical discovery software from SAS Institute, Inc. See FIG. 5 , agraph of Kaplan-Meier (product-limit) survival estimates showing thefraction of bees surviving over time in days when given extracts of themycelium of Fomes fomentarius (0.1%, 1%, and 10% as respectively shownby dotted, dashed and double-dash lines) with sugar water as compared toa control population fed sugar water only (shown by a solid line). Thisanalysis compared the mean survival time for the treatment to the meansurvival time for the control (fed just sugar water) and used a Wilcoxontest to assess whether the survival was statistically different fromchance variation in bee survival time. FIG. 5 illustrates thisembodiment of the invention. In this analysis a composition of Fomesfomentarius fed at 1% v/v improved the mean survival of bees by 9.7%(p<0.0006) over the duration of the test. Notably however, is that therewas a near doubling of the number of bees surviving the first week, atime that correlates to peek pollen gathering activity by worker bees,with extracts of Fomes fomentarius. A composition of Fomitopsis pinicolamycelium extract fed at 0.1% v/v (volume/volume) showed a similarimprovement the number of worker bees within the last three weeks oftheir lives.

As demonstrated in FIG. 1-5 , longevity of bees fed extracts ofdifferent fungal species was improved, with improvement in longevitydependent on both the fungal species and the concentration consumed bythe bees. In the practice of the current invention, some of thisincrease in longevity is probably due to a reduction in viral burden inmost instances (as discussed below), but is also attributable to otheraspects of the invention in the instance where longevity improved butviruses were not reduced.

Improvement in longevity can be demonstrated in the practice of theinvention by the use of survival plots, such as but not limited to,those described above. Improvement in longevity can be measurednumerically in the practice of the invention by calculating thedifference in survival. One such method is based on the average value ofa function theorem:

$f_{avg} = {\frac{1}{b - a}{\overset{b}{\int\limits_{a}}{{f(x)}{dx}}}}$Where values of ‘a’ and ‘b’ represent the starting and ending days overwhich, the effect of the invention is being measured, and ‘f(x)’represents the survival plot function as previously described. Thedifference in these figures over the specified time interval representsaverage percent improvement in longevity achieved through the practiceof the invention over the specified time interval.

Other methods for measuring differences in longevity, survival, orpopulation increases, including statistical methods such as Kaplan-Meyeranalysis, Nelson Aalen, and other methods for which are known to thoseskilled in the art are acceptable alternatives in the practice of theinvention.

Similarly, various quantitative methods of assaying virus numbers inbees may be utilized in the practice of the present invention, includingreverse transcription Polymerase Chain Reaction (RT-PCR) and real-timeRT-PCR based on the PCR amplification of cDNA, ELISA (enzyme-linkedimmunosorbent assay), including both normal and sandwich ELISA with thevarious blocking agents, primary/secondary antibodies, reporter enzymesand their specific colorimetric substrate solutions for detection andquantification, multiplexing microarrays utilizing molecular probes fordifferent target RNAs or DNAs, AGID (Agarose Gel Immuno-diffusion),serology methods based on protein profiles or polyclonal and monoclonalantibodies and the large variety of other molecular biology basedmethods such as high throughput sequencing technologies,pyrophosphate-based sequencing techniques, Sanger sequencing (alsoreferred to as the chain termination method) and integrated virusdetection systems (IVDS). See, for example, De Miranda, Diagnostictechniques for virus detection in honey bees, in Aubert et al. (Eds.),Virology and the honey bee, EEC Publications (2008), pp. 121-232 andEvans et al., Standard methodologies for molecular research in Apismllifera, Journal of Apicultural Research 52(4) (2013).

Using this method for measuring the difference in longevity, theinventor specifies the improvement in longevity as embodied by thisinvention. See Table II, “Average Percent Improvement in Longevity ofBees.” The table represents the difference between the average values of% of bees surviving, when assessed over various time intervals. Thatdifference is given as the numerical subtraction of these percentages,with the average percent surviving over various time intervalscalculated as previously described:longevity improvement=avg % surviving_(fed fungal extract)−avg %surviving_(control)Improved longevity increases the number of “bee days” in which workersor other classes of bees are available to gather pollen and maintain thehive or perform other labor, whereby the improved health and increasedsurvival of the individuals leads to improved colony health andsurvival.Reduction in Total Virus Level

For antiviral testing of each type of mycelium extract (mushroomspecies), mixed aged honey bees from a single hive were collected on asingle day and distributed at random into four cages of roughly 100 beeseach. This trial was done in parallel to the longevity testingpreviously described, using bees from the same hives over the same timeinterval. Each fungal species set consisted of a control cage (fed sugarsyrup), a low concentration cage (fed mycelium extract in sugar syrup at0.1% v/v), a medium concentration cage (fed mycelium extract in sugarsyrup at 1% v/v), and a high concentration cage (fed mycelium extract insugar syrup at 10% v/v).

Samples of bees were removed from the cage and were frozen at day 0, day7, and day 14. Assay of the total number of virus particles,irrespective of viral species, was carried out by Dr. David Wick of BVS,Inc. utilizing IVDS technology; see U.S. Pat. Nos. 8,524,155, 8,309,029,8,146,446, 8,021,884, 7,850,908, 7,250,138, 6,491,872, 6,485,686 and6,051,189 (all to Charles Wick) and Charles H. Wick, Integrated VirusDetection, CRC Press (2014). For each sample analysis, 6.0 grams of beeswere blended with 100 ml of Reverse Osmosis (RO) water and coarsefiltered through dual layer cheesecloth. A 90 ml sub-sample was thencentrifuged for 60 minutes at 20,000×g. The supernatant was recoveredand ultrafiltered through a 500,000 Dalton hollow fiber filtrationsystem followed by a rinse with a 200 ml RO wash and reduction toapproximately 2 mL. The solution was prepared for Integrated VirusDetection System (IVDS) analysis using a 1:10 dilution with AmmoniumAcetate (AA). Each sample was filtered through a W-41 20 μm paper or a0.45 μm PTFE filter. Samples were scanned five times with the IVDS, andaverage virus levels were reported.

As demonstrated in FIGS. 6, 7, 8 and 9 , the total viral load of beesfed extracts of different fungal species was reduced, with the level ofvirus reduction dependent on both the fungal species and theconcentration consumed by the bees. FIG. 6 is a bar graph showing totalvirus particles in bees given extracts of the mycelium of Inonotusobliquus (0.1%, 1%, and 10% as respectively shown by dotted, dashed anddouble-dash lines) with sugar water as compared to a control populationfed sugar water only (shown by a solid line) at time zero, one week andtwo weeks. FIG. 7 is a bar graph showing total virus particles in beesgiven extracts of the mycelium of Ganoderma resinaceum (0.1%, 1%, and10% as respectively shown by dotted, dashed and double-dash lines) withsugar water as compared to a control population fed sugar water only(shown by a solid line) at time zero, one week and two weeks. FIG. 8 isa bar graph showing total virus particles in bees given extracts of themycelium of Fomitopsis pinicola (0.1%, 1%, and 10% as respectively shownby dotted, dashed and double-dash lines) with sugar water as compared toa control population fed sugar water only (shown by a solid line) attime zero and one week. FIG. 9 is a bar graph showing total virusparticles in bees given extracts of the mycelium of Schizophyllumcommune (0.1%, 1%, and 10% as respectively shown by dotted, dashed anddouble-dash lines) with sugar water as compared to a control populationfed sugar water only (shown by a solid line) at time zero and two weeks.These and similar figures can be used in the practice of the inventionto demonstrate the effect of the invention on the health of the bees. Inthe practice of the current invention, most but not all, of the speciesthat had improved longevity also had reduced virus load. This impliesthat viral reduction can help longevity; but that improvements inlongevity may be seen without viral reduction because of otherbeneficial aspects of the invention such as general stimulation of hiveimmunity and antibiotic activity against non-viral pathogens likeNosema. Multiple causes of longevity improvement are likely in thepractice of the invention because different fungal species appear havedifferent and specific modes of action against different bee pathogensas disclosed below, by way of example and not exclusion.

Reduction in total viral load can be measured in the practice of theinvention by calculating the difference in the virus detection betweenbees to which the invention has been applied and bees which have notbeen exposed to the invention. One such method for quantifying thisdifference is based on the average value of a function theorem:

$f_{avg} = {\frac{1}{b - a}{\overset{b}{\int\limits_{a}}{{f(x)}{dx}}}}$Where values of ‘a’ and ‘b’ represent the starting and ending days overwhich, the effect of the invention is being measured, and ‘f(x)’represents the virus detection level as a function of temporal sampling.The “percent difference” in these values over the specified timeinterval represents average “percent reduction” in virus level achievedthrough the practice of the invention over the specified time interval.Other methods for measuring differences virus level over time, includingpercent difference at individual sampling time points, mean difference,and statistical methods such as Kaplan-Meyer analysis are acceptablealternatives in the practice of the invention and are incorporated byreference. Using the method described above for measuring the differencein virus level over various time intervals, the inventor specifies thereduction in virus as embodied by this invention. See Table I, “AveragePercent Decrease in Total Viral Burden.”

The table represents the difference between the average values of % ofbees surviving, when assessed over various time intervals. Thatdifference is given not as the numerical subtraction of thesepercentages, but rather as the “percent reduction”:

${\%{decrease}{in}{viral}{burden}} = {\frac{{{avg}{virus}{titer}_{{fed}{fungal}{extract}}} - {{avg}{virus}{titer}_{control}}}{{avg}{virus}{titer}_{control}} \times 100}$Fungal Species/Disease Specificity

Specific types of mycelium extract (mushroom species), when fed to mixedaged bees can reduce the level of disease-causing agents such as virusparticles in a species-specific way. This embodiment of the inventionwas demonstrated by feeding fungal extracts to caged bees and measuringthe levels of specific types of virus in the bees over time. For thisanalysis, mixed aged bees from a single hive were collected on a singleday and were evenly distributed at random into 12 cages. Four cages werefed Ganoderma resinaceum mycelium extract at 1% v/v in sugar syrup (50%sucrose; 50% water; weight/volume, w/v, which, with water, is equivalentto weight/weight or w/w), four cages were fed Inonotus obliquus myceliumextract at 1% (volume/volume, or v/v) in sugar syrup, and four were usedas a control and were fed only sugar syrup.

Samples of bees were removed from the cage and were frozen at day 0, day3, day 7 and day 14. Bees were sent to Dr. Yanping (Judy) Chen of theUnited States Department of Agriculture—Agricultural Research Serviceusing real time RT-PCR as described in Chen et al., Quantitativereal-time reverse transcription-PCR analysis of Deformed Wing Virusinfection in the honeybee (Apis mllifera L.), Appl. Environ. Microbiol.,Vol. 71 (2005), p. 436-441 and Khongphinitbunjong et al., Differentialviral levels and immune gene expression in three stocks of Apis mlliferainduced by different numbers of Varroa destructor, Journal of InsectPhysiology, Vol. 72 (2015), p. 28-34.

In this analysis, levels of virus are quantified based on theaccumulation of a fluorescent signal as DNA of the virus is amplified inthe PCR reaction. The cycle threshold is defined as the number of cyclesrequired for the fluorescent signal to exceed the fluorescencebackground level. Cycle threshold levels are therefore inverselyproportional to the amount of target viral nucleic acid (e.g., virustiter) in the sample (i.e., the lower the CT level the greater theamount of target nucleic acid in the sample). As demonstrated in FIG. 10, the levels of Black Queen Cell Virus (as quantified based on cyclethreshold) were prevented from increasing in bees that were fed extractsof Ganoderma resinaceum and Ionotus obliquus mycelium. See FIG. 10 , aline graph showing cycle threshold for Black Queen Cell Virus over timein a control population fed sugar water only (shown by solid line 111)as compared to bees given extracts of the mycelium of Inonotus obliquus(1%) (shown by dotted line 112) and Ganoderma resinaceum (1%) (shown bydashed line 113) with sugar water.

This example of a fungal species extract having specificity against oneviral pathogen and not against another embodies the antiviral invention.It also supports the argument that specific compositions of fungalextracts can be similarly specific to other bee pathogens (that reducelongevity) such as Nosema and bacteria and/or can, in general,up-regulate metabolic, immune and detoxification systems of bees. Sucheffects against non-viral pathogens or general metabolic and immunityboosts may have been responsible for the instances where longevity wasimproved but viral load remained unchanged.

Summary, Preferences, and Implications

To date, the inventor has data (both a longevity experiment and a totalvirus reduction experiment) for 8 species of medicinal mushrooms usingmycelium grown on rice. Overall viral reduction is reported below, aswell as longevity metrics. The inventor and his team have, incollaboration with WSU and the USDA, have recently conducted additionalexperiments to test against specific viruses, using extracts of myceliumgrown on rice and extracts from mycelium grown on sawdust. The specificviruses tested showing activity include the Deformed Wing Virus, theBlack Queen Cell virus, and the Lake Sinai virus.

The inventor herein defines a metric, the “LV index,” which is: LVIndex=The average percent improvement in bee longevity multiplied by theaverage percent decrease in total viral burden.

This computation gives a number that assigns equal importance to bothaspects for measuring improvement to colony health. Blank boxes in thetables below for antiviral activity, longevity or LV indicate thateither longevity or virus reduction was negative or zero in one or bothdata sets.

There are many other possible mathematical representations that coulddraw a relationship between these data sets, such as, for example,percent longevity improvement divided by percent virus reduction. Thatcalculation would stress the portion of the longevity that couldtheoretically be related to virus reduction if there were a 1:1correspondence between these measures. Numerous possibilities for ametric will be apparent to those skilled in the arts and all suchmetrics for improved bee health should be considered to be within thescope of the invention.

The general approach adopted herein is to compare the “area under thecurve” of longevity measurements and total virus reduction measurementsas previously described. The difference between the areas under thecurves, over a given time interval, is equal to the numbers in thelongevity table. The difference in the area under the curves, over agiven time interval, expressed as a “percent improvement” is equal tothe numbers in the total virus reduction tables. See Table I “AveragePercent Decrease in Total Viral Burden” and Table II “Average PercentImprovement in Longevity of Bees.” These values can then be relatedmathematically to illustrate features of interest in the practice of theinvention such as compositions that are most preferred for improvinglongevity and reducing total viral load in bees. See Table III AveragePercent Improvement in Longevity of Bees×Average Percent Decrease inTotal Viral Burden (LV Index).

TABLE I Average Percent Decrease in Total Viral Burden ConcentrationSpecies Timeframe 0.1% 1% 10% Trametes versicolor 0-7 days 7.5 9.0 0-14days 4.8 3.6 Fomitopsis pinicola 0-7 days 20.4 22.8 0-14 days 25.5 32.22.1 Fomitopsis officinalis 0-7 days 0-14 days 4.5 Schizophyllum commune0-7 days 3.8 0-14 days 19.5 20.8 26.7 Inonotus obliquus 0-7 days 47.541.6 42.2 0-14 days Fomes fomentarius 0-7 days 9.6 10.0 0-14 days 9.3Ganoderma applanatum 0-7 days 2.5 3.6 1.0 0-14 days 4.5 14.2 Ganodermaresinaceum 0-7 days 73.4 64.4 76.7 0-14 days 65.4 58.3 85.9 Preferred1-25% or greater decrease in virus More Preferred 15-25% or greaterdecrease in virus Most Preferred >25% decrease in virus

TABLE II Average Percent Improvement in Longevity of Bees ConcentrationSpecies Timeframe 0.1% 1% 10% Trametes versicolor 0-7 days 4.1 0.2 5.00-14 days 3.8 7.8 0-28 days 1.6 3.7 Fomitopsis pinicola 0-7 days 9.214.6 0-14 days 13.5 14.1 0-28 days 8.5 3.7 Fomitopsis officinalis 0-7days 5.2 3.8 0-14 days 2.3 1.0 0-28 days 1.0 Schizophyllum commune 0-7days 0.1 0.8 0-14 days 3.7 0.5 0-28 days 0.1 Inonotus obliquus 0-7 days1.6 1.2 0-14 days 4.1 2.1 0-28 days 3.7 Fomes fomentarius 0-7 days 1.722.1 13.5 0-14 days 16.1 0-28 days 11.2 Ganoderma resinaceum 0-7 days0-14 days 2.2 3.7 0-28 days 3.7 9.5 Preferred 1-5% or greaterimprovement in longevity More Preferred 3-5% or greater improvement inlongevity Most Preferred >5% improvement in longevity

TABLE III Average Percent Improvement in Longevity of Bees × AveragePercent Decrease in Total Viral Burden (LV Index) Concentration SpeciesTimeframe 0.1% 1% 10% Trametes versicolor 0-7 days 30.9 44.8 0-14 days18.1 27.7 Fomitopsis pinicola 0-7 days 187.5 332.7 0-14 days 344.1 453.1Fomitopsis officinalis 0-7 days 0-14 days 10.4 Schizophyllum commune 0-7days 3.1 0-14 days 72.6 9.6 Inonotus obliquus 0-7 days 67.0 51.2 0-14days Fomes fomentarius 0-7 days 16.2 135.2 0-14 days Ganodermaapplanatum 0-7 days 0-14 days Ganoderma resinaceum 0-7 days 0-14 days147.1 212.9 Preferred 1-200+ LV index More Preferred 50-200+ LV indexMost Preferred 200+ LV index

In the practice of the invention, fungal extract compositions may bevariously ranked with regard to preference depending on the intendedapplication of the composition. Examples include but are not limited toranking with preference to longevity improvement, ranking withpreference to total virus reduction, ranking with regard to longevityand virus reduction. Notably, preference may also be given to methodsand compositions, which improve longevity but do not reduce viruses.Such compositions are expected to improve longevity by acting on beestressors that are unrelated to viruses (examples include Nosemainfection, pesticide exposure, stress from cold temperatures, etc.).

The process of mixing mycelial extracts with sugar, water, and optionalingredients (such as those in pollen patties) and feeding to bees ispreferred.

Example 9

Ganoderma resinaceum extract at 14 days resulted in an almost 20%increase in survival of worker bees over the controls. See also FIG. 5 .This differential can be hugely significant in helping the colonysurvive as the longevity of worker bees during this critical timeresults in nurse bees not being prematurely recruited, thus allowingthem to better attend to keeping the brood, the next generation,healthy. The addition of mycelial extracts from Ganoderma resinaceumresulted in a dramatic reduction in overall viral pathogen payloads inbees (from multiple viruses), while the sugar control, without mycelialextracts, resulted in increased population of overall viruses. Asviruses are thought by many bee entomologists to be the most significantdisease challenge, often facilitating the subsequent infection fromother bacterial (i.e., foulbrood) and fungal species, reducing virusescan be a keystone advantage in protecting bees from colony collapsedisorders and their many associated stressors.

In terms of increasing longevity, the addition of 1% mycelial extractsof Fomes fomentarius and Ganoderma resinaceum to sugar water (water-50grams, sugar 49.5 grams, mycelial extract 0.5 grams), statistically,significantly extended the lifespans of bees—in terms of tee days oflife' by 17.6% and 8.9%, respectively. Extended average lifespan resultsin more workers being available for job tasks, a significant advantageto stressed bee colonies on very thin operating margins and stressedcolonies on the edge of collapse. When there are more bees at any onetime that is significant for pollen acquisition and hive maintenance. Byextension, many more hives can be saved feeding them mycelial extractsin their sugar water over those just having sugar water without mycelialextracts. Until field trials, it is unknown how many more bee days willtilt the balance to help bees overcome CCD since there are so manycomplexities. However, the consensus amongst bee scientists is thatincreasing longevity of worker bees, under stress, is a strongadvantage. Moreover, when the extracts are made from, in these cases,birch tree wood (Betula species), the same tree species these polyporemushrooms habit, and ones in which bees nest, the extracts may becomemore potent while less expensive to produce. That we can show suchstrong, significant activity from mycelium grown on both rice andsawdust strengthens the argument that the mycelium is the causalbenefitting factor (the rice controls showed no activity.) By utilizingmycelium grown on rice as spawn to inoculate 10-100× more mass in theform of birch sawdust expands the mycelium exponentially over themycelium-on-rice extracts reported here. The mycelium grows more denselybranched and compacted mycelial networks on birch sawdust compared torice, meaning more surface areas is generated for the expression ofextracellular constituents. Hence, mycelial extracts from birch or otherwood sawdust will be a preferred embodiment of this invention.

Many Ganoderma and other polypore species are anticipated to also offera similar ‘bioshield’ of protection. No doubt, there will be gilledmushrooms, due to their close evolutionary relationship to polypores, tobe of benefit similarly.

Example 10

The inventor's Amadou Fomes fomentarius and Red Reishi Ganodermaresinaceum extracts were prepared from mycelium grown on birch sawdustfor approximately two months. These “1×” extracts were evaporated infront of a HEPA filtered laminar flow hood for several days, resultingin an approximate 10:1 reduction, and removing ˜95% of the residualethanol and ˜90% of water, creating what we termed “10×” extracts. These10× extracts, in the form of a viscous syrup, were then fed at a 0.1%and a 1% concentration in the sugar-feed water of caged honey bees forabout three weeks at Washington State University under the supervisionof Steve Sheppard and Brandon Hopkins. Data was collected via qRT-PCRfrom pooled 30-50 bees analyzed by Jay Evans at the ARS/USDA.

In analyzing the antiviral test results of a 1% extract concentration ofFomes fomentarius (“F.f.”) and Ganoderma resinaceum (“G.r.”) myceliagrown on birch sawdust vs. extracts of the birch sawdust controls, thereduction of the Deformed Wing Virus (DWV) and the Lake Sinai Virus(LSV) is as follows:

Fomes Fomentarius Extract from Birch vs Bee DWV:

F.f. vs. DWV 1024:1   Birch Control vs DWV 128:1  F.f. vs LSV 8:1 BirchControl vs LSV 4:1Ganoderma Resinaceum Extract from Birch vs Bee DWV:

G.r. vs. DWV 64:1 Birch Control vs DWV  8:1 G.r. vs LSV 512:1  BirchControl vs LSV 32:1It will be noted that, unlike rice controls, the birch controls showanti-viral activity. The inventor notes sawdust comes from trees, andvirtually all trees host fungal populations, to varying degrees. Using afungal free sawdust control is problematic and unrealistic. In doing DNAanalysis of the birch sawdust controls, these three fungal species wereamplified to a level of significance of detection: Graphostroma sp.(probably G. platystoma), Chondrostereum purpureum and Trametesversicolor.

Graphostroma platystoma is an ascomycetes (Ascomycota) of the OrderXylariales whereas Chondrostereum purpureum and Trametes versicolor arebasidiomycetes (Basidiomycota) of the order Polyporales. Hence, theinventor anticipates, as a derivative of his invention, that species ofascomycetes and additional basidiomycetes will be reservoirs hostingantiviral properties. By controlling the fungal species growing undercontrolled clean-room laboratory conditions, the dominant fungi will bethat which is selected, producing thousands of more times the fungaltissue than any native fungi within freshly milled sawdust.

WSU oversaw tests of extracts derived from mycelium grown on sterilizedrice using Chaga, Inonotus obliquus (“I.o.”), and Red Reishi, Ganodermaresinaceum (“G.r.”), which showed greater than 500× reductions of theBlack Queen Cell Virus (BQCV).

Inonotus obliquus Extract from Myceliated Rice vs Vee BQCV:

I.o. vs. BQCV >500:1Ganoderma Resinaceum Eextract from Myceliated Rice vs vee DWV:

G.r. vs. BQCV >500:1

The process of cultivating a mushroom mycelium on wood or sawdust,extracting the mycelium, and mixing mycelial extracts with sugar, water,and optional ingredients (such as those in pollen patties) and feedingto bees is preferred.

Example 11

In coordination with Washington State University and under thesupervision of Dr. Steve Sheppard, Chair of Entomology, outdoor fieldtrials were conducted in mid-September 2016 in Idaho with beehiveshosting queens for testing ETOH/H₂O extracts made from myceliated birchsawdust colonized by Fomes fomentarius and Ganoderma resinaceum againstviruses. Commonly known as ‘nucs’ by the bee industry, these small5-frame beehives contained colonies composed of about 8000 worker beesand a queen.

Ten beehives each were treated with Fomes fomentarius and Ganodermaresinaceum mycelial extracts from 64 and 62 day, respectively, incubatedcultures grown on sterilized birch sawdust as two separate sets with oneset of 10 beehives as a control. Control and treatment colonies were fedsucrose syrup (50% sucrose; 50% water, by volume) in internal framefeeders.

The hydroethanolic mycelial extracts (1×) were added at a concentrationof 1% of total volume, or 10 mL per 1000 mL (1 liter) to their sucrosesyrup feed water. (This is ˜ 1/10 of the concentration listed in Example10. ) Bees were fed and consumed 3 liters of the extract-enriched feedin less than a week. Some beehives consumed the feed water supplementedextracts by Day 4, after which time additional sucrose syrup (withoutextracts) was provided. On Day 7, a second treatment of 3 liters ofcontrol syrup or syrup containing mycelium-based extracts was provided.On Day 4, Day 15, and Day 21 bees were sampled and frozen for latervirus analysis. Samples of bees were sent and tested for viral titers byDr. Jay Evans and his team, of the United States Department ofAgriculture (USDA-ARS Bee Research Lab BARC-E Bldg 306 Beltsville, Md.20705 USA).

Dr. Evans and the USDA analyzed samples and provided a summary of theantiviral effects of a 1% addition of ETOH/H₂O extracts from myceliatedbirch sawdust dissolved into the sugar syrup. Two feedings were made attime 0 and on day 7.

On Day 15:

Compared to controls, Fomes fomentarius and Ganoderma resinaceum birchsawdust extracts reduced the Deformed Wing Virus by >8:1.

Compared to controls, after incubating for 62 and 64 days respectively,Ganoderma resinaceum birch sawdust extracts reduced the Lake Sinai Virusby >500:1 and Fomes fomentarius birch sawdust extracts reduced the LakeSinai Virus by >32:1.

Example 12

A liquid extract of the mycelium, or a precipitate from such extract, ora concentrated extract from which all or part of the solvent has beenremoved, containing these active principles can be added to the honey,to honey-enriched water, to sugar water or bee candy, to pollen, topollen substitutes, or to other substances in other manners obvious tothose skilled in the art of apiary science or commercial practices. Theextract can be used as an adjunct to other remedies making them moreeffective. The extracts can be in liquid, frozen, freeze dried, airdried, vacuum desiccated, refractance window dehydrated, sonicallydehydrated, or partially purified forms, in amounts sufficient to havethe effect of attracting bees and/or benefitting bee health, honeyproduction and pollinations. Moreover, these derivative forms ofextracts will be useful for human consumption as they are palatable,high in antioxidants, and in other properties beneficial to people andother animals, including bees.

Example 13

Unique combinations of extracts from the metabolites from differentspecies of polypore fungi can be created to afford the greatestprotection from a plethora of bee damaging viruses. For instance,hydroethanolic extracts Fomitopsis pinicola mycelium grown on sterilizedrice can be combined with hydroethanolic extracts of Fomes fomentariusgrown on sawdust. Fomitopsis pinicola extracts extend longevity whilstFomes fomentarius extracts reduce viruses. Combining them both augmentthe bee-longevity effects of either alone. Moreover, these mixtures canbecome much more complex, to target a plurality of viruses andstressors. For instance, a mixture of an extract of Fomes fomentariuswhich reduces Deformed Wing Viruses, with an extract of Inonotusobliquus which reduces the Black Queen Cell Viruses, with Ganodermaresinaceum which reduces the Lake Sinai Viruses, would give beekeepersthe convenience of protecting and preventing viral infection before thebeekeeper's detection of what virus was about to devastate theirbeehives. Adding longevity-extending extracts such as Fomitopsispinicola will further enhance the usefulness of this complex and uniquemixture. Many other species are expected to provide additional benefits.By periodically changing the combinations of the fungi included in thesemixtures, acquired viral resistance to these treatments is less likely.Hence, this inventor is actively engaged in surveying populations ofwood decomposing fungi to optimize suites of formulas for long termusefulness for protecting and extending longevity of bees. Since thesewoodland mushroom species grow throughout many forests of the world,local strains of fungi can be isolated from natural habitats using themethods described in the author's patents and books. Moreover, somepolypore mushrooms are resupinate, meaning they do not erect a fleshyfruitbody, but form a crust, like Irpex lacteus. The inventoranticipates these crust polypore mushrooms will now be candidates worthyof testing for bee and animal benefits.

Example 14

A preferred delivery system uses means for incorporating the mycelialextracts into pollen patties or grease patties. Pollen patties are madeby beekeepers and placed above the brood chamber as a source ofnutrition. They can be made from a wide range of materials, includingsoy, brewer's yeast, sugar syrup and may optionally include organicallygrown pollen. These pollen patties supplement the bees nutritionally.Because they are widely used in the fall, they help the bees surviveinto the next year. These extracts also contain digestive enzymes whichhelp the bees better metabolize food stocks and help break down toxinsand improve baseline immunity. The process of incorporating or mycelialextracts into pollen patties or grease patties via mixing withappropriate ingredients and forming the patties and presenting to beesis preferred.

Example 15

A mixture of compositions comprising extracts of Strophariarugosoannulata, Fomes fomentarius, Fomitopsis officinalis, Fomitopsispinicola, Ganoderma resinaceum, Inonotus obliquus, Piptoporus betulinus,Trametes versicolor and/or Schizophyllum commune, which together offer aplurality of benefits, can be added to water. The Strophariarugosoannulata attracts bees, has a flower-like fragrance, and providessugar-rich (up to 75 polysaccharides) nutrient source. The Fomesfomentarius and Fomitopsis officinalis extracts confer antiviralbenefits, plus those additional benefits already mentioned forStropharia rugosoannulata. All three extracts contain polyphenols, andmore particularly coumaric acids and coumarins, some of which helpactivate p450 enzyme pathways, which help bees detoxify endogenous,natural, foreign, and anthropogenic toxins and their associateddeleterious effects. A mixture of these extracts can be given to thebees via their drinking water, their enriched water, honey, propolis,pollen patties or even in the wax used for making preformed combs in thecreation of supers for honey production.

Example 16

Add the extracts from the mycelium of Fomitopsis officinalis, Fomitopsispinicola, Fomes fomentarius, Inonotus obliquus, Schizophyllum commune,Ganoderma resinaceum, Piptoporus betulinus, Trametes versicolor and/orInonotus obliquus to the sugar-water typically fed to bees in the earlyspring before pollen levels rise, to help reduce resident viral loadsearly in the season, preventing their escalation to the level ofbecoming a behavior-altering disease or for causing bee-to-bee transferof pathogens. The extracts can simply be mixed into the sugar water at arate sufficient to have a positive effect. The range could preferably be0.01-20%, or more preferably 0.1-10% of the volume of the sugar watercompositions employed by beekeepers. The extracts would be mixed in thewater first and then added to the sugar to make the typical syrup. Onestandard formula would be to add 0.1 to 1% of the extract v/v to a sugarsyrup feeding solution.

The following formula and working example indicate preferred ranges forextract and bee feeding supplement solutions or non-miticidal beesprays. (The percentages for liquids are volume/volume or v/v. Thepercentage of dry ingredients is weight/volume or w/v):

-   -   0.1-20% mushroom mycelium extract in any form from any means of        extraction;    -   2-67% glucose, maltose, sucrose, or fructose (or other sugars or        sugar enriched complexes, where sugar >50%);    -   33-50% water by weight to weight, w/w;    -   0-0.1% sodium benzoate, EDTA or other preservative in an        effective amount or 2-200 ppm chlorine bleach (‘chlorinated        sugar syrup’); and    -   0-10% glycerol or polyethylene glycol, or other thickeners,        wetting agents, surfactants and surface active agents,        dispersants, emulsifiers, solubilizing agents, tackifiers or        adhesives, penetrants, carriers, antibiotics or nutritional        supplements, dispersants, humectants, arrestants, feeding        stimulants, sex pheromones, aggregating pheromones, vanillic        acid, semisolid cellulosic, hemicellulosic, lignocellulosic, and        lignin substrates untreated or treated with chitinases,        amylases, or other enzymes.

The following formula and working example indicate preferred ranges formiticidal bee sprays:

-   -   0.1-20% (v/v) mushroom mycelium extract in any form from any        means of extraction;    -   2-50% (w/v) glucose, maltose, sucrose, or fructose (or other        sugars or sugar enriched complexes, where sugar >50%);    -   2-50% water;    -   then by w/v 2.5-4.2% anhydrous oxalic acid dihydrate or 3.5-5.0%        oxalic acid dihydrate (or other miticides in an effective        amount); and    -   0-0.1% sodium benzoate, EDTA or other preservative in an        effective amount or 2-200 ppm chlorine bleach.

Example 17

Preferred liquid sprays include aqueous solutions, emulsifiableconcentrates, emulsions such as oil-in-water and water-in-oil emulsions,dispersions, suspoemulsions, microemulsions, water-dispersible granules,wettable powders, microcapsules, etc. Wettable powders are formulationsthat are typically uniformly dispersible in water and also containsurface active agents (surfactants) such as wetting agents, emulsifiers,and dispersing agents. Emulsifiable concentrates are prepared withorganic solvents and/or one or more emulsifiers. Sticking agents such asoils, gelatin, gums, tackifiers and adhesives may be used to improve theadhesion of the spray. Humectants may also be used to decrease the rateof evaporation, including for example glycols having from 3 to 10 carbonatoms and glycerin and solutes such as salts or sugars in water.

In another preferred embodiment, mycelial extract is added to sprayhydroseeding equipment, mobile landscaping hydroseeders, or foggers.Spray hydroseeding is performed with a pump for dense liquids, whichsprays on to the surface to be treated a mixture consisting of, forexample, mycelial extracts and optional fungal inocula (spores, driedhyphae, powdered mushrooms, conidia, etc.), fertilizer if desired orsoil improvement substances, optionally and usually preferably with abinder or tackifiers, and water.

For many landscaping and agricultural applications, use of cart-mountedor trailer-mounted hydroseeding or fogging units and the mobilevariations will be preferable. Hydroseeding units are typically utilizedto plant lawn grasses, and may be utilized to plant native grasses,wildflowers, mixtures of grasses, shrubs, bushes, trees, crops, etc. ifdesired. Spores, fresh mycelium, dried or freeze-dried mycelium,powdered mushroom fruitbodies, the many forms of fungi imperfecti andtheir conidia (asexually produced spores) and related fungal forms andcombinations thereof may be easily added to the hydroseeding mixture.Hydroseeding units typically employ mechanical agitation (via paddles oraugers inside the tank) or jet mixing (via pump jets) of water andmaterials; other methods will be readily apparent to those skilled inthe art.

Another preferred embodiment of the present invention is the use ofextracts with agricultural equipment, including planting equipment,harvesting equipment, field preparation equipment and processingequipment with means for delivering extracts. Appropriate methods ofmodifying agricultural equipment with pumps, sprayers and/or mixers,etc. will be readily apparent to those skilled in the art. Spores,mycelial hyphae and or powdered mushrooms may optionally be introducedinto agricultural equipment via methods known to the art so as toprovide the benefits of simultaneous inoculation with saprophytic,endophytic, mycorrhizal or entomopathogenic and/or other beneficialfungi.

By way of example but not of limitation, such agricultural plantingequipment may include foggers, seeders, air seeders, planters, airplanters, plate planters, vacuum planters, drills, air drills, airseeding systems, row crop cultivators, planting systems, inter-row orbetween row planting systems, rice transplanters, etc.

Agricultural harvesting equipment may include, by way of example only,combines, round balers, square balers, hay cubers, threshers andthreshing machines, forage harvesters, windrowers, rakes, tedders,mowers, rotary mowers, sicklebar mowers, slashers and cutters, strawchoppers, stalk choppers, corn pickers, cotton strippers and gins, cornhuskers, shellers, rice harvesters, mechanical fruit and nut pickers,loaders, etc.

In another preferred embodiment, such extracts may be utilized directlywith agricultural equipment useful for preparation and/or improvement offields, orchards, etc. Such equipment includes by way of examplesprayers, foggers, irrigators, plows, cultivators, air carts, fertilizerspin spreaders, pendulum spreaders, etc.

Agricultural areas including, for example, cranberries bogs, blueberrypatches and almond orchards may be treated with means for sprayingextracts as a foliar treatment. By spraying extracts of, for instance,Fomes fomentarius mycelium, dual benefit protecting plants and bees canbe realized with one application. Fomes fomentarius extracts on blueberries, or on cherry, apple or almond orchards, even on alfalfa orrapeseed canola crops, as a few examples of many, could both reducemosaic (many of which are Ilarviruses) viruses that harm plants whilsthelp native and honey bees better survive from the deleterious effectsof the Deformed Wing Virus and/or other viruses. This two-for-onesolution using one species of fungus to address two classes of virusesis, to the best of this inventor's knowledge, unprecedented. Thisinvention now allows for formulating mixtures of antiviral fungi can beconcocted to give a broad host defense of resistance of viruses thatharm plants and animals sharing habitats or food chains. The beneficialagricultural implications are staggering to contemplate. For instance,just for the fruit tree and nut agricultural industries, the need forsolutions for alleviating the damage from mosaic viruses could save manybillions of dollars by improving crop yields:

-   -   “These studies, coordinated by the Mediterranean Agronomic        Institute of Bari (Italy), involved 14 countries of Southern and        Eastern Europe, Middle East, and North Africa, representing        areas with different germplasm. The results of enzyme-linked        immunosorbent assay (ELISA) testing of approximately 24,000        trees (almond, apricot, plum, peach, and sweet and sour cherry)        demonstrated a high incidence (23.5%) of ilarvirus infection. Of        those infected trees, mixed infections were recorded in 76.4% of        the trees. The ilarvirus infection was distributed as follows:        PNRSV (46.4%), PDV (40.7%), ApMV (3.3%), and mixed infection,        mostly PNRSV and PDV (9.6%). Among the different stone fruit        species, cherry was the most infected (45.6%), followed by        almond (24.5%), peach (24.4%), plum (15.2%), and apricot (6.2%).        Prevailing single viruses in different species were PDV in        cherry (35.4% of the total tested or 71% of the infected trees)        and PNRSV in peach (17% of the total tested or 53.3% of the        infected trees).”        Pallas, V., Aparicio, F., Herranz, M. C., Amari, K.,        Sanchez-Pina, M. A., Myrta, A., and Sanchez-Navarro, J. A. 2012.        Ilarviruses of Prunus spp.: A continued concern for fruit trees.        Phytopathology 102:1108-1120.

Example 18

The antiviral extracts described in this invention can be diluted intowater and used as a foliar spray utilizing all technologies and meansfor spraying designed for foliar applications in agriculture or for pestcontrol. Moreover, the antifreezing foggers commonly used by the citrusindustry could emit clouds of antiviral droplets whose direct contactwith the plants could expose not only bees but many other insects to theantiviral benefits of the inventor's extracts. Thus, this invention isanticipated to reduce many viruses afflicting plants besides mosaicviruses. And since this inventor has noted extracts from Ganodermalucidum and Ganoderma resinaceum resist freezing (not freezing whenbelow <10° F.), the foliar spray made from these extracts mayadditionally protect plants and crops from damaging freezing temperatureexposure.

Example 19

Fungally Treated Structural Materials to Help Protect Bee Colonies

Beehives are typically made of wood fibers, and stackable ‘supers’ holdframes upon which bees create honeycombs rich in honey, above the broodboxes. These supers and the other wood-based frameworks used bybeekeepers eventually degrade, often times growing molds that can bepathogenic to bees and encourage invasion by insects and arthropods. Asa result, beekeepers typically replace their supers, brood boxes, andfloors etc. every few years. The bases of beehives tend to last longer,but ultimately all beehive boxes degrade with opportunistic, oftenpathogenic fungi, and the health of bee colonies can be negativelyimpacted as a result.

This invention is to add, incorporate, impregnate, coat, infuse orattach spores or fragments of mycelium, or extracts thereof, ofbeneficial fungi, more specifically mushroom forming fungi in the ClassBasidiomycetes/Basidiomycota or Ascomycetes, into the frameworks ofbeehives, which impart a benefit to bee health by providing a hostdefense shield of singular or synergistic benefits. For instance, andnot by limitation, the frames of supers can be made of sheets of wood,wood fiber, and a wide range agricultural products than be boundtogether with mycelium, with or without spores. Such fungallyimpregnated panels not only provide a structural benefit, but the fungican have antiviral, anti-miticidal, antibacterial, antifungal,anti-insecticidal properties that help bees dwelling on and within thematerials used for building beehives.

In addition to beehives, all structures that house animals may beadvantageously treated with extracts or mycelium, including bird cages,barns, and structures to house bats.

The materials that can be used to host these beneficial fungi can bemade of wood, particle boards, biodegradable structured panels, fungallygrown wood panel substitutes, non-biodegradable materials, and may alsobe incorporated into the bees wax used for the frames, or even thepropolis bees use for closing gaps. Compositions and methods of thisinvention can easily be incorporated into such open source beehivemanufacturing.

In essence, as the beehives age, they incrementally, sporadically, orincreasingly provide a health benefit to bees from the addition of theabove-mentioned fungi to offset the increased stressors that occur withbeehives boxes as they get older. The antiviral properties of thesefungi, their creation of a wide assortment of complex and simple sugars,the production of p-coumaric acid and other cytochrome p450 activatingmolecules, the formation of spores of entomopathogenic fungi likeMetarhizium anisopliae that harm mites but not bees, all worksynergistically to counteract the myriad stressors bees suffer fromduring the lifespan of colonies in hives year to year.

One of numerous examples would be to grow mycelium of, for instance, thepolypores Antrodia cinnamonea, Fomes fomentarius, Inonotus obliquus,Inonotus pini, Fomitopsis officinalis, Fomitopsis pinicola, Fomitiporiarobusta, Ganoderma applanatum, Ganoderma australe, Ganoderma atrum,Ganoderma annulare, Ganoderma annularis, Ganoderma brownii, Ganodermalingzhi, Ganoderma lucidum, Ganoderma resinaceum, Ganoderma tsugae,Irpex lacteus, Heterobasidion annosum, Lenzites betulina, Phellinusigniarius, Phellinus pini, Phellinus weirii, Piptoporus betulinus,Trametes elegans, Trametes versicolor and the gilled mushroomsSchizophyllum commune, Stropharia rugosoannulata and Lenzitesponderosus, Psilocybe cubensis on agricultural waste materials so thatthe mycelium can be formed into the structural panels used to build beeboxes for honey bees. Additionally, the mite-killing fungus Metarhiziumanisopliae, can be grown in a preconidial (pre-sporulating) or postconidial (post sporulating) state—or a mixture thereof—and be embeddeddirectly into the constructed panels, no matter what their composition,in combination with the fungi described herein as well as other speciesthis inventor anticipates that could help bees in the Basidiomycetes,Ascomycetes, or the Entomophthorales, Hypocreales, or other fungiattacking mites or phorid flies.

Moreover, the aforementioned species and their many relatives containand secrete toxin-degrading enzymes which can help break downinsecticides (including neonicotinoids, neonics), fungicides,pesticides, formaldehydes, tannins, dyes, endemic toxins, and a widearray of anthropogenic toxins.

A preferred wood-based substrate would be composed of birch panels,boards, or sawdust (Betula species) and some preferred species would bethe birch polypores such as, but not limited to Fomes fomentarius,Inonotus obliquus, Ganoderma resinaceum and Piptoporus betulinus. Ifmade of conifer woods, then polypores such as Fomitopsis pinicola,Fomitopsis officinalis, Laetiporus suiphureus, Phellinus igniarius, P.pini, P. linteus and P. weirii; or gilled mushrooms such Pleurotusostreatus, Lentinus ponderosus are ideal candidates. Trametes versicolor(=Coriolus versicolor) is a polypore mushroom growing on deciduous andconifer woods and is also a preferred species to deploy within thecontext of this invention. With each of the species listed, they are tobe considered in the broadest concept of the species, i.e., ‘sensulato,’ and close relatives are also anticipated to be useful to helpingbees. As such, when describing Fomitopsis officinalis, Ganodermaapplanatum, Ganoderma lucidum, Ganoderma resinaceum, Inonotus obliquus,Trametes versicolor, or any other mushroom species, this meansFomitopsis officinalis sensu lato, Ganoderma applanatum sensu lato,Ganoderma lucidum sensu lato, Ganoderma resinaceum sensu lato, Inonotusobliquus sensu lato, Trametes versicolorsensu lato and a similar broaddescription of any other species, each of which means that this is thespecies concept as described within the broadest taxonomicinterpretation, encompassing all historical and modern synonyms,varieties, forms and species that have or will be split from thesespecies since publication. As is known in the art, names change as newspecies concepts are constructed. The species anticipated to be usefulis extremely broad, many of which have been listed in the inventor'spreviously approved 8 U.S. patents and within the pending patentapplications filed to date. Nevertheless, those species not previouslylisted now become obvious, subsequent to this inventor's discovery.

That the polypore mushrooms Fomes fomentarius, Inonotus obliquus andGanoderma resinaceum are active against viruses that harm bees andhumans is remarkable, and to the best of the knowledge of this inventoris, medically, unprecedented. Moreover, if these cross-animal benefitscan be obtained from the mycelial extracts of these polypore mushrooms,and indeed many mushrooms, then more than one animal species may benefitfrom the vast antiviral properties from the mycelia of these species.Hence, bird houses, chicken houses, barns and animal housings of anysort, bird feeders and plant trellises may be constructed of cellulosicproducts with the addition of these cultures, their extracts or theirspores for immunological and community-protection benefit, preventingdisease vectors from escalating and even curing illnesses of itsresidents within. Potentially homes using mycelium and fungi couldprotect residents from viruses, bacteria, insects, arthropods, toxins,environmental stressors, disease vectors, and unexpectedly impartpleasant fragrances specific to the fungi deployed.

Extracts useful for the above invention can come as a by-product ofthose using mycelium for filling forms or molds to create mycelium grownstructured materials, such as insulation, shipping materials to replaceStyrofoam, building materials, packaging materials, filtration cushions,filtration membranes, fabrics, scaffolding for growing mycelial basedcomputer chips and processors, mycobacterial based nanowires, etc.Additionally, these useful extracts can be harvested by expressing theliquid components from substrates used in all stages of mushroomproduction as well as from the fungal fermentation methods used formaking tempeh, koji, enzymes, antibiotics, plant growth enhancers, andpharmaceuticals. In essence when growing out the mycelium, themycelially made materials often are dried. In doing so, theextracellular and intracellular metabolites and other liquids must beremoved. When growing of mycelium based structured materials, thisexcess liquid is discarded and not typically highly valued. Thisinvention repurposes this ‘waste’ liquid product into an unexpected highvalue-added suite of products that can be rich in antivirals,antimicrobials, enzymes, acids, active ingredients, and other chemicalsuseful to this invention for helping bees and for many otherapplications in medicines, chemical engineering, degradation practices,and bioremediation (mycoremediation). Moreover, the now dried myceliatedproduct can be designed so that a latent population of fungal cellssurvive the drying process, only to be re-activated when the bee hivesage, causing the mycelium and its heat-tolerate sclerotia andchlamydospores to survive and re-grow to provide an unusual benefit—asthe bee hives age, the impregnated beneficial fungi compete againstfungal pathogens, provide nutrients, increase overall bee colonylongevity. Beneficial fungi can be selected specifically for heatresistant chlamydospores and sclerotia survivability subsequent to themanufacturing of mycelially grown, structured materials. The repurposedliquid from compressing the mycelium as well as the heat-tolerantmycelium resident within the structured materials can be combined forsynergistic benefits to bee health.

Example 20

Although many beekeepers feed their honey bees sugar water, andantiviral, longevity enhancing mycelial extracts as described herein canbe easily deployed for helping domesticated honey bees, native wild beesare at a disadvantage as they cannot easily gain access due tocompetition at the honey bee hive. Putting extract enriched sugar waterinto hummingbird feeders seems like an obvious choice but fail due tocompetition from other insects, in particular yellow jackets and wasps.Yellow jackets are notorious for their invasion of beehives and arearchrivals to bees as are many wasps.

By adapting existing hummingbird or gerbil feeders, this inventorenvisions a solution.

A standard feeder can have two sections for feeding: one for bees andone for bee competitors.

Utilizing a device having two or more sugar water emitters, as used forhummingbirds, gerbils, or for other creatures, these ports for feedingbees can be physically separated from the ports feeding bee competitors.One elaboration is to have a dividing wall splitting the feeder in half.The wall facing the ports feeding the bees would be bluish in color asbees are highly sensitive and can see far deeper in the blue spectrumthan most insects. The bluish colors are highly attractive to bees. Thewall facing the ports feed bee competitors would be yellow, or of anon-blue color more attractive to the bee competitors than to the bees.The dividing wall would separate the competing species who would ratherfeed that fight, if ample food is apportioned, and territoriallyseparated from one another.

As noted in this patent, attractiveness by bees to blue light, which isinvisible to humans but visible to bees, is a highly significantdiscovery as bees are most easily trained to associate food in theultraviolet wavelengths of color. As Menzel and Backhaus determined in1989, bees could learn faster when the food was associated with violetlight was used compared to all other colors. Menzel, R. and Backhaus, W.1989. “Color vision in honey bees: Phenomena and physiologicalmechanisms.” In D. Stavenga and R. Hardie (eds.): Facets of vision.Berlin-Heidelberg-New York: 281-297.

A combination of blue spectra, even using static or pulsing LED UVlights, that could be optionally battery or solar powered, could aid inthe bees quickly learning which side to feed from. Using sensors andwireless transmitters, data collection can be enabled which would notonly chart the presence and feeding behavior patterns, but would alertvia text, email, or website, the time to replenish the feeders.Moreover, the extract enriched sugar water can have dyes or any coloringagent that will give the extract enriched feed water a bluish color,that could further augment attractancy and discovery by bees. Suchdevices can be elaborated upon in many ways and improved withexperience. Sonic wave emitters and electromagnetic transmitters canalso be added to this invention to further improve their function.

Example 21

Culture the medicinal mushroom mycelium on plant materials that haveactivity against viruses, including Ficus bengalensis (Vad), Ficusreligiosa (Pimpal), Jasminum auriculatum (Jaai), Acacia catechu (Khair),Azadirachta indica (Neem), Curcuma longa (Turmeric), Withania somnifera(Ashwagandha) and Silybum marianum (Milk Thistle). See Deshpande et al.,Antiviral activity of plant extracts against sac brood virus in vitro—apreliminary report, International Journal of Institutional Pharmacy andLife Sciences 3(6): November-December 2013, p. 1-22. Extract utilizingmeans for extracting and means for treating bees utilizing means forfeeding bees or means for spraying bees or other means for drenching orother means for wetting, dousing, or bathing bees are preferred.

Example 22

Use extracts of the mycelium or fruitbodies from Ganoderma lucidum,Ganoderma resinaceum, Fomitopsis pinicola, Fomitopsis officinalis,Inonotus obliquus, Piptoporus betulinus, Trametes versicolor andSchizophyllum commune whereby the extracts are concentrated into a formattractive to bees and sufficient, upon contact, to have the effect ofreducing the Tobacco Ringspot Virus, the Israeli Acute Paralysis Virus,the Black Queen Cell Virus, the Invertebrate Iridescent Virus, or IIV6,and Nosema microsporidia, resulting in bees being able to betterovercome colony collapse disorder.

Use extracts of the mycelium or fruitbodies from Ganoderma lucidum,Ganoderma resinaceum, Fomes fomentarius, Fomitopsis pinicola, Fomitopsisofficinalis, Schizophyllum commune, Inonotus obliquus and Strophariarugosoannulata whereby the extracts are concentrated into a form thatresembles the texture and consistency of honey, in a form attractive tobees and sufficient, upon contact, to have the effect of reducingviruses, including but not limited to the Tobacco Ringspot Virus, theIsraeli Acute Paralysis Virus, the Black Queen Cell Virus, and Nosemamicrosporidia, and causing the up-regulation of cytochrome p450 enzymepathways, improving overall immune function, foraging ability,overwintering, drought resistance, ability to overcome losses of nectarproviding plants, resulting in an improved health to bees so that thereis a measurable benefit for beehives to survive and overcome CCD andproduce descendent generations. This “mycological honey” can be usedseparately or mixed into bee honey to attract and benefit bees.Moreover, this “mycological honey” can be partially dissolved into wateras a foliar spray to plants or applied directly onto bees. Additionally,this ‘mycological honey’ can be marketed as a nutraceutical for humanconsumption.

Example 23

Use extracts of the mycelium or fruitbodies lacking melanin such as fromso called albino fruitbodies of Agaricus blazei, Fomitopsis officinalis,Fomitopsis pinicola, Fomes fomentarius, Schizophyllum commune, Trameteselegans and Stropharia rugosoannulata whereby the extracts areconcentrated into a form that resembles the texture and consistency ofhoney, in a form attractive to bees and sufficient, upon contact, tohave the effect of reducing the Tobacco Ringspot Virus, the IsraeliAcute Paralysis Virus, Invertebrate Iridescent Virus, or IIV6, andNosema microsporidia, and causing the up-regulation of cytochrome p450enzyme pathways, improving overall immune function, foraging ability,overwintering, drought resistance, ability to overcome losses of nectarproviding plants, resulting in an improved health to bees so that thereis a measurable benefit for beehives to survive and overcome colonycollapse disorder and produce descendent generations. This “mycologicalhoney” can be used separately or mixed into bee honey to attract andbenefit bees. Moreover, this “mycological honey” can be partiallydissolved into water as a foliar spray to plants or applied directlyonto bees. Additionally, this ‘mycological honey’ can be marketed as anutraceutical for human consumption.

Example 24

Extracts of medicinal mushroom mycelium can be soaked into paper stripsand other cellulosic cloths or fabrics. These can be combined with anadhesive. The low pH of the many medicinal mushroom mycelium extracts,in the pH 0.5-4 range, is toxic to mites but harmless to bees uponcontact. Oxalic acid and formic acid solutions may optionally be addedin effective amounts.

Example 25

Bees flying to or from the sugar water, upon entering the beehive, buzzand shake their bodies to dislodge the mites. If the mites fall througha screen, they are in contact with or attracted to the entomopathogenicmycoattractant, which in itself may be lethal, or onto insecticidalmycelium, wherein the mites sicken or die, reducing the mites' abilityto travel and infect, thus lessening its threat vector to bees.Moreover, if bees are sprayed with an oxalic acid enriched spray, theparasitic mites become more susceptible to the infectious or lethalproperties of the entomopathogenic fungi. The processes of cultivating amushroom mycelium on grain or sawdust, extracting the mycelium andmixing mycelial extracts with water, a miticide and optional ingredientssuch as sugar, and spraying bees, beehives, beehive components, areassurrounding beehives and areas to be pollinated by bees are preferred.

Example 26

The extracts, hyphal fragments or spores of beneficial fungi, such asFomes fomentarius, and the spores of entomopathogenic fungi such asEntomophthorales, can be incorporated as a mixture into theextract-enriched sugar water, bee foods, honey or sprays, which allowsfor transference into the honey production stream, benefitting thebrood, the drones, the queen and the hive overall.

Example 27

Extracts of the mycelium of, or spores, hyphal fragments, or tissue of,Stropharia rugosoannulata can be presented on paper strips or in wateraccessible to the bees. The fragrance of Stropharia rugosoannulata, towhich bees can be accustomed, helps foraging bees to return to theircolonies if these fragrances are placed near to or within the hives.Such fragrances can be emitted via any method known to the art ofdelivery of fragrances, foggers, sprays, or aerosol dispensers. It isexpected that the extracts of Stropharia rugosoannulata mycelium and theextracts of other mushroom mycelia will induce trail following ornavigation behavior via “dance language” and odor plumes.

Example 28

Spores and hyphae of Metarhizium anisopliae may be mixed with theextracts or dried forms made from the mycelium of Fomes fomentarius forproducing anti-Varroa mite sprays and smokes for helping bees resistmites, viruses, etc. for overcoming CCD. Many strains of Metarhizium arerelatively nontoxic; “No harm is expected to humans from exposure toMetarhizium anisopliae strain F52 by ingesting, inhaling, or touchingproducts containing this active ingredient.” Metarhizium anisopliaestrain F52 (029056) Biopesticide Fact Sheet. The spores of Metarhiziumanisopliae can be immersed into glycol, mineral oil, or sugar syrupsalong with extracts or dried forms of antiviral benefitting mushroomspecies described within this patent such as Fomes fomentarius,Ganoderma resinaceum and Inonotus obliquus. A suggested concentration ofspores (conidia) of Metarhizium anisopliae or other entomopathogenicfungi such as Beauveria bassiana would be >100,000-1,000,000 permilliliter in the final aliquot mixture.

Example 29

A mixture of compositions of extracts of Stropharia rugosoannulata,Fomitopsis officinalis, Fomitopsis pinicola, Fomes fomentarius,Ganoderma resinaceum, Inonotus obliquus, Piptoporus betulinus, Trametesversicolor and/or Schizophyllum commune and Metarhizium anisopliae,which together offer a plurality of benefits, can be added to water. TheStropharia rugosoannulata attracts bees, has a flower-like fragrance,and provides sugar rich (up to 75 polysaccharides) nutrient source. Thevarious extracts confer antiviral and antibacterial benefits and lifeextension, plus the attractancy of Stropharia rugosoannulata. TheMetarhizium anisopliae extracts can be presented in sticky strips ormats, or into any sticky, mite- or Phorid fly-capturing substance, or inwater accessible to the same to attract mites and Phorid flies,whereupon contact, they are debilitated or killed, reducing theirability to be a vector of disease; Varroa mite populations can bereduced using Metarhizium anisopliae extracts before the brood chambersare sealed, reducing bee deaths from exposure to mites and the diseasesthey carry. All three extracts contain polyphenols, and moreparticularly coumaric acids, which help activate p450 enzyme pathways,which help bees detoxify endogenous, foreign, natural, and anthropogenictoxins and lessen their associated deleterious effects. A solution ofthese mixed extracts can be given to the bees via nectar feeders, insideor outside the hives, containing their drinking water or their sugar orfructose enriched water, via mixing into bee candy, honey, propolis,pollen patties or even by mixing into the wax used for making preformedcombs in the creation of supers for honey production.

Example 30

Extracts of the preconidial mycelium of Metarhizium anisopliae and/orBeauveria bassiana pathogenic to mites and/or flies can be mixed withspores or hyphal fragments of same, and presented in sticky strips ormats, or into any sticky, mite- or Phorid fly-capturing substance, or inwater accessible to the mites. This combination attracts mites or flies,which upon contact, infects them with an entomopathogenic fungus orexposes them to a lethal doses of entomopathogenic toxins.

Example 31

Extracts of the preconidial mycelium of Metarhizium anisopliae mixedwith the extracts, spores, or hyphal fragments of Strophariarugosoannulata can be presented on paper strips or in water accessibleto the bees. This combination attracts mites or flies, and bees, whichupon contact harms the mites and flies but not bees.

Example 32

Extracts of the preconidial mycelium of Aspergillus flavus, Aspergillusniger and Aspergillus fumigatus can be mixed with the spores or hyphalfragments of Stropharia rugosoannulata and presented on paper strips orin water accessible to the bees. This combination attracts mites orflies, and bees, which upon contact harms the mites and flies but notbees. Optionally, strains of Aspergillus flavus, Aspergillus niger andAspergillus fumigatus can be used which have reduced aflatoxin andneurotoxin levels, below the levels which would harm bees but above thelevels harming mites and flies, thus conferring a net benefit to beecolony health.

Example 33

Extracts of the preconidial mycelium of Metarhizium anisopliae can bemixed with the spores or hyphal fragments of Stropharia rugosoannulatacan be presented on paper strips or in water accessible to the bees.This combination attracts mites or flies, and bees, which upon contactharms the mites and flies but not bees. Optionally, strains ofMetarhizium anisopliae can be used which have reduced destructin levels,below the levels which would harm bees but above the levels harmingmites and flies, thus conferring a net benefit to bee colony health.

Example 34

Extracts of mushroom mycelium and/or extracts of the preconidialmycelium of Metarhizium anisopliae can be mixed with extracts orderivatives from Neem trees and presented on paper strips, in wateraccessible to the bees or in topical sprays. This combination attractsmites or flies, and bees, which upon contact harms the mites and fliesbut not bees. Optionally, strains of Metarhizium anisopliae can be usedwhich have reduced destructin levels, below the levels which would harmbees but above the levels harming mites and flies, thus conferring a netbenefit to bee colony health. Optionally, the concentration of Neem treeextracts (or the active ingredient azadirachtin), and sugars can bebalanced to optimize benefits to bees by reducing mites and theirforaging abilities, and their pathogen payloads. Furthermore, thiscombination can be further enhanced with the addition of extracts ofBasidiomycetes fungi from agaricoid and polyporoid fungi, which not onlyprovide mite-destroying oxalic acids, and toxin degrading enzymes, butalso up-regulates bee's innate cytochrome p450 enzymatic pathways tobreak down anthropomorphic toxins, and additionally reduces virally,bacterially, and fungally associated pathogens afflicting bees. Suchsynergistic effects from multiple constituents have the net effect ofhelping bees better survive colony collapse disorder. A combination ofusing preconidial mycelium of Metarhizium anisopliae, the extracts ofFomitopsis officinalis and Fomitopsis pinicola, the extracts from Neemtrees, the extracts of Ganoderma lucidum, Ganoderma resinaceum,Ganoderma applanatum, Pleurotus ostreatus, Trametes versicolor andStropharia rugosoannulata immersed and mixed into water is anticipatedto be an effective composition and method for making a deliverable,efficacious bee spray or ingredient in pollen patties or drinking water.Similar compositions may be sprayed on plants or trees which beespollinate, benefitting both plant and bee.

Example 35

The methods and compositions of oxalic acid, sugar (or polysaccharide)enriched water, and the preconidial hyphal fragments from Metarhiziumanisopliae which upon contact with bees selectively harms the miteswhile having a net benefit to bees. This composition may optionally becombined with extracts of medicinal mushroom mycelium with antiviral andlongevity enhancing properties and incorporated into bee food and beesprays.

Example 36

The combination of the extracts from Fomes fomentarius, Fomitopsispinicola, Fomitopsis officinalis, Ganoderma lucidum, Ganodermaapplanatum, Ganoderma resinaceum, Piptoporus betulinus, Schizophyllumcommune, Stropharia rugosoannulata and Ganoderma resinaceum incombination with the extracts of the preconidial mycelium of Metarhiziumanisopliae to attract bees and mites whereby contact with thiscombination harms Varrroa mites, reducing viruses, pathogenic fungi andbacteria, providing a net benefit for bees overcoming colony collapsedisorder.

Example 37

The combination of the preconidial mycelium of Metarhizium anisopliaewith polysaccharides of Fomitopsis pinicola, Fomitopsis officinalisGanoderma resinaceum, Ganoderma lucidum, Inonotus obliquus, Piptoporusbetulinus and Schizophyllum communes, to treat bees and mites wherebycontact with this combination harms Varrroa mites, and reduces viruses,bacteriophages, pathogenic fungi and bacteria that harm bees but has anet benefit for bees overcoming colony collapse disorder.

Example 38

Extracts of the spores, mycelium and hyphal fragments, or fruitbodytissue of the polypore mushrooms Fomes fomentarius, Fomitopsis pinicola,Fomitopsis officinalis, Piptoporus betulinus, Ganoderma resinaceum,Ganoderma lucidum, Schizophyllum commune, and Inonotus obliquus, can beaerosolized, or delivered via droplet-clouds, sprayed into hives incombination with a spore-mycelium mixture of Metarhizium anisopliae.Such a mixture will reduce viral pathogens, up-regulate detoxificationpathways within bees, provide a wide assortment of complex and simplesugars, vitamins to augment immunity and longevity of the hive.Moreover, the addition of Metarhizium anisopliae spores and mycelium caninfect, control, and prevent Varrroa mites and Phorid flies. Theendogenous oxalic acid within all these fungi can help control thesepathogens. The individual use or combinations of the birch polyporesfungi can restrict growth of the Nosema microsporidium fungi, andfoulbrood bacteria (such as the bacterium Melissococcus plutonius) frominflicting harm to the bee colony. Such aerosolized sprays can bedelivered using handheld or backpack sprayers. Alternatively, absorbentstrips can be deployed infused with the above beneficial agents.Constructed materials used for building the housings of bee boxes canalso be soaked and impregnated with these beneficial agents.

Example 39

α-Amylase, amyloglucosidase, betulinic acid, caffeic acid,protocatechuic acid, trans-cinnamic acid, ferulic acid, gallic acid,ellagic acid, lanosterol, inotodiol, trametenolic acids, hispolons,ergosterols, chrysin, cordycepin, trans-o-coumaric acid,trans-p-coumaric acid, ellagic acid dihydrate, ergosterol, linoleicacids, trans-ferulic acid, gallic acid hydrate, hexanal, hispolon,4-hydroxybenzoic acid, quercetin hydrate, rutin hydrate, syringic acid,vanillic acid, sulpherinic acid, dehydrosulphurenic acid, eburicoicacid, 6-chloro-4-phenyl-2H-chromen-2-one, ethyl6-chloro-2-oxo-4-phenyl-2H-chromen-3-carboxylate,7-chloro-4-phenyl-2H-chromen-2-one, ethyl7-chloro-2-oxo-4-phenyl-2H-chromen-3-carboxylate, psilocybin, psilocinand their congeners, isomers, structural analogs and significantlysimilar compounds may prove useful in the practice of this invention.The compounds are also anticipated to be useful with other animals,including humans. Vanillic acid, hispolon, quercetin hydrate, rutinhydrate, syringic acid, trans-cinnamic acid, trans-ferulic acid andsalts of vanillic acid, syringic acid, trans-cinnamic acid andtrans-ferulic acid and ethyl7-chloro-2-oxo-4-phenyl-2H-chromen-3-carboxylate are preferred for theirantiviral activity; see U.S. patent application Ser. No. 14/853,932.

Since protocatechuic acid, vanillic acid, cinnamic acid, ferulic acid,caffeic acid and their congeners, isomers, structural analogs andsignificantly similar compounds are widely distributed and present inmany edible plants and with protocatechuic acid being naturally high inbrown rice bran and grain, growing bee-benefitting fungal speciesmentioned herein on a substrate already containing protocatechuic acid,its precursors and analogs, will likely increase the net amount of thesebee-benefitting compounds, and is a preferred embodiment.

Attractants:

One component of the invention is the use of fungal extracts, wherebythe extracts are generated from the mycelium of polyporoid,basidiomycetous and ascomycetous species, to attract bees. The bees areattracted to the polysaccharide-rich extracellular and intracellularmetabolites secreted by the mycelium. Within these exudates arecompounds that attract bees, feed them with sugar rich and othernutrients, provide antiviral, antifungal, and antibacterial protection,while bolstering their resistance to pesticides and improving colonyhealth and honey production. In fact, honeys holding these fungalcomponents could proffer medicinal benefits to bees and other animalspecies, including humans.

These extracellular exudates from, for instance, the King Stropharia orthe Garden Giant mushroom (Stropharia rugosoannulata), have anattractive effect on bees, especially during the time when floweringplants of their preference are limited. Bees are attracted both to theextracellular extracts as well as living mycelium. Other non-toxicmushroom species, which may or may not possess antiviral and lifeextending properties, including gourmet and medicinal mushrooms, areexpected to attract bees to varying degrees in a similar fashion.

The pleasant fragrance of Stropharia rugosoannulata out-gassing from themycelium may attract bees, although no other scientists, to the best ofthis inventor's knowledge, has ever discovered or stated this. Thepresent inventor has discovered Stropharia rugosoannulata mycelium emitsa rich, attracting flower-like essence. Oyster mushrooms in the genusPleurotus, especially Pleurotus ostreatus, P. pulmonarius, P.lignatilis, P. sapidus, P. eryngii, P. populinus and other relatedspecies emit a pleasing anise-like fragrance, as does Clitocybe odora.Another candidate is the split-gill polypore, Schizophyllum commune, oneof the most common of all woodland Basidiomycetes, which produces apotent, sweet fragrance in culture, at times overwhelming the olfactorysenses of lab personnel, and is a source of coumarins and coumaricacids. Interestingly, only those growing Schizophyllum commune in mass,in vitro, on cereal grains or wood would ever know about this potentoutgassing fragrance. The inventor knows of no one else in his 40 yearsof experience who has mentioned or reported on this fragrance phenomenonwith this species. Schizophyllum commune is one of the most prominentwhite rot, woodland species across the temperate and tropical regions ofthe world, and creates softened, sweet wood from which bees can benefit.Many other species probably emit attractive fragrances to bees, whichare undetectable to humans or not noticeably enticing.

The mycelium from Agaricomycetes and the extracts made from the pureculture mycelium may be the source of new bee attractants. TheAgaricomycetes are the only fungi that decompose lignin, and includesthe gilled mushrooms, such as Stropharia rugosoannulata, and thepolypores, such as those related to Fomitopsis species. TheAgaricomycetes encompasses ˜16,000 described species. Many of theAgaricomycetes dually decompose cellulose and lignin. Native bees userotten logs for nesting, as discussed above in connection with bears,fungi, and bees, which the inventor hypothesizes provides bees with thesugar rich and cytochrome P450 coding and up-regulating compounds viawater droplets and nectar secreted by the mycelium of Agaricomycetes.

Currently, our regenerated forests have about 10-15% of the wood debriscompared to native woodlands! This relatively recent loss ofdecomposable wood debris limits the availability of these beneficialfungi to native and imported bees, introducing a heretofore unreported,additional stress factor. The continued constriction of debris fieldsfurther erodes the food webs essential not only to bees, but also tomost organisms that are dependent upon healthy and sustainableecosystems.

For instance, fungal extracts of the preconidial (pre-sporulation)mycelium of non-Agaricomycetes fungi, including Metarhizium anisopliaeand Aspergillus flavus, have been shown by the inventor to attractPhorid flies and other insects, arresting their migration, and thusprevent these flies from vectoring diseases. See U.S. Pat. Nos.6,660,290; 7,122,176; 7,951,388; 7,951,389; and 8,501,207. Moreover,pathogen hosting mites are also attracted and stopped from moving intothe bee colonies using these mycelium-based extracts, thus reducing notonly the pathogen payloads mites carry, but also reducing the numbers ofmites which might otherwise infect the bees. Similar approaches may beused to control beehive pests, such as the greater and lesser wax mothsand the small hive beetle, if needed. Moreover, strains of thesepre-sporulation entomopathogenic fungi can be selected for their highthermal tolerance and their abilities for attracting and killing mitesand flies which harm bees or vector pathogens. Research intopost-sporulation and spore-based Metarhizium anisopliae technologies(which may have the disadvantage of repelling mites and/or insects ascompared to the attractancy of preconidial mycelium) have demonstratedthe relative ease with which strains may be selected for thermaltolerance to high hive temperatures and high pathogenicity and/ormortality to Varrroa mites. Rodriguez et al., Selection ofentomopathogenic fungi to control Varroa destructor (Acari: Varroidae),Chilean J. Agric. Res., 69(4): 534-540 (2009); Rodriguez et al.,Evaluation of Metarhizium anisopliae var. anisopliae Qu-M845 isolate tocontrol Varroa destructor (Acari: Varroidae) in laboratory and fieldtrials, Chilean J. Agric. Res., 69(4): 541-547 (2009); Boyle, NewBrunswick Department of Agriculture, Aquaculture and Fisheries,Integrated Pest Management-Compatible Biological Control of Varroa Miteof Honey Bee; Fungi help combat honeybee killer, BBC NewsScience/Nature, Aug. 9, 2002.

Moreover, the inventor has clearly shown that the preconidial myceliumof entomopathogenic fungi, such as but not limited to, Metarhiziumanisopliae, Beauveria bassiana and Cordyceps species, elicit astimulatory feeding response (phagostimulation) in many insects andother arthropods from the smelling and subsequent ingestion of theextracts made from presporulating (preconidial) mycelium. However, beesshow a unique tolerance to the toxins from the spores and mycelium ofMetarhizium anisopliae that harms mites and phorid flies. Hence having ablend of entomopathogenic fungi, prior to sporulating, or extractsthereof, mixed with the spores (conidia) of these same fungi, couldstimulate the bees to consume more mycelia and the extracts thereof,including the beneficial polypore fungi, resulting in a unique suite ofsynergistic advantages, which includes longevity factors, antiviral,antibacterial and antifungal effects, up-regulations of cytochrome(p450) detoxification pathways, providing complex sugars, vitamins andnutrients, while lessening the toxicity of anthropogenic insecticides,herbicides, fungicides, anthropogenic toxins and also reducing mite andphorid fly populations, all the while introducing fungal speciessupporting a healthy bee gut bacterial microbiome. Each one of thesefactors helps bees reduce the stressors of colony collapse disorders.The combination of these benefits within one delivery system—as acomposition or a method—is an unprecedented approach, to the best of theknowledge of this inventor. Methods for selecting and optimizing strainswithin each species will likely result in improvements as each variableis tested and combined.

Humans are limited to perceiving color wavelengths of light fromapproximately 390 to 750 nanometers (nm). Bees, like many insects, seecolors from approximately 300 to 650 nm. Many mushroom species likeOyster mushrooms (Pleurotus ostreatus) are triggered into fruitingaround 360 nanometers, beyond the far end of our ability to detect. (SeeAction spectra for Hyphal Aggregation, the first stage of fruiting, inthe basidiomycete Pleurotus ostreatus, Richartz and Maclellan inPhotochemistry and Photobiology pages 815-820, May 1987. Mushroommycelium will absorb some of this light and reflect much of it, due tothe limitations of absorption through the translucent, hyaline cellwalls of the mycelium.

When mycelium growing deep within wood or the ground reaches the surfaceof ground or wood, and is exposed to light, a phase change occurs in themushroom's life cycle, going from mycelium to the first stages ofmushroom formation, hyphal aggregation and primordia ('baby mushroom')formation. The mycelium in many species will not form primordia unlessthere is light exposure near to the ultraviolet or 360 nanometer orlower wavelengths. This is well within the range bees can detect butbeyond the limits of what humans can.

Attractiveness to mycelium stimulated by blue light invisible to humansbut visible to bees is highly significant discovery as bees are mosteasily trained to associate food in the ultraviolet wavelengths ofcolor. As Menzel and Backhaus determined in 1989, bees could learnfaster when the food was associated with violet light was used comparedto all other colors. Menzel, R. and Backhaus, W. 1989. “Color vision inhoney bees: Phenomena and physiological mechanisms.” In D. Stavenga andR. Hardie (eds.): Facets of vision. Berlin-Heidelberg-New York: 281-297.

Hence, bees finding surfacing mycelium, at the time when nutrients arebeing up-channeled into the pre-primordia or primordia forming myceliumin response to violet light wavelengths, and when this light is criticalfor stimulating mycelium to switch into mushroom formation, suchdetection by bees would be an opportune time to find surfacing myceliumand capture dense nutrition when mycelium is so metabolically active.Although hypothetical and speculative by this inventor, this interactionmerits further research since bees can be trained to discover food basedon light spectra associations. This added element to this invention canaccelerate the learning process of bees for finding new food sourcesusing the attributes of mycelium. As a result, the embodiments of thisinvention also provide the benefit of enhancing the usefulness andattractiveness of other forms of foods for helping the health of beesusing these aforementioned mycelial properties, particularly helpingbees discover mycelium at the primordia formation stages.

Surfacing mycelium outgasses carbon dioxide and exudates fragrances, andthis inventor hypothesizes that bees can detect mycelium not only fromits scent, but are also attracted to the mycelium's response to thisblue spectrum light, whereupon mushroom mycelium begins to pack protein,vitamins, and sugar-rich nutrients at the interface between the highcarbon dioxide environment within substrates and the highly oxygenatedenvironments just above, and in doing so builds nutritionally dense butaccessible primordia—the first stage of mushroom formation orbasidiospores formation (as in the case of resupinate polypores likeInonotus species, forming exposed hymenial surfaces, or crusts, that arebrightly colored such as Inonotus andersonii). Many of the brightlycolored fungal pigments, especially but not limited to yellowish ones,exhibited by mycelium can be composed of fungal bioflavonoids, many ofwhich are polyphenols. Exploring this rich interface environment—thesurface of yellowish fungal mycelial membranes exposed to theatmosphere—is anticipated by the inventor to be a rich reservoir forbees to harvest extracellular and intracellular metabolites endowed withnutrients and immune-supporting compounds, including “mycoflavonoids”and “mycosterols” including phenols and polyphenols not limited tocoumarins and benzoic and cinnamic acid derivatives including coumaricacids and their glycosides.

By way of example, but not of limitation, mycelia of some species,especially in the genus Phellinus and Inonotus, produce brightlycolored, yellowish pigments in their mycelium including polyphenols, forexample hispolons such as6-(3,4-dihydroxyphenyl)-4-hydroxyhexa-3,5-dien-2-one, (C₁₂H₁₂O₄), abright yellow bioactive group of compounds with antioxidant and immuneenhancing properties derived from polypore species such as Inonotushispidus and Phellinus linteus. The inventor hypothesizes these brightyellowish-colored mycelia would additionally attract bees foraging forsugars, polyphenols, moisture, natural nutrients, and other secretionsthat have immune-building antiviral, antibacterial, antifungal, andantiprotozoal properties. Since bees are especially attracted to yellowcolors, those species of fungi, such as Phellinus and Inonotus, whichproduce bright yellowish colors, could preferentially attract bees andalso are directly associated with the yellowish polyphenols containingcoumarins to help bees activate their cytochrome P450 enzyme pathways.This inventor sees the growing of these wood-decomposing species thatproduce brightly pigmented mycelia as preferred candidates for designingmycelial platforms and extracts for helping bees. Consequently, extractsof mycelium forming primordia and extracts of colored mycelium arepreferred bee attractants.

Moreover, sawdust extracts may prove useful for antiviral applicationsother than bees. The combinations of these fungi, grown on sterilizedsawdust can provide p-coumaric acid to activate the cytochrome p450pathways needed by bees for detoxifying xenobiotic chemicals. Byexposing the mycelium to light at specific wavelengths, especially inthe yellow and blue wavelengths, antiviral or virostatic molecules canbe triggered into production. Thus, extracts of this mycelium provide amultiplicity of benefits whose combinations are unique and unprecedentedin the history of science for humans helping bees. Fungal species andthe type of substrate (sawdust, grain, etc.) may be combined to afford aunique mixture helping bees, insects, and other animals, and likelyprovide plants protection against pathogenic viruses.

Integrative Fungal Solutions for Protecting Bees

The first antiviral from a mushroom ever discovered was from the “IceMan” polypore, Fomes fomentarius, against the Tobacco Mosaic Virus, thefirst virus ever to be discovered, and related to the Tobacco RingspotVirus, and now within a large clade of mosaic viruses. This polyporemushroom is a saprophyte on birch, beech, and other temperate deciduoushardwoods. When it grows, the wood is softened, releasing moisture,insect-attracting fragrances and sweetened with the rich, complexpolysaccharides, as well as proteins and other substances generated bythe mycelium of this fungus. This fungus attracts beetles, which becomefood for woodpeckers and other birds, whose excavated burrowssubsequently can be occupied by native bees. In essence, this is oneexample of what the inventor anticipates being many examples of the rolepolypore and other Basidiomycetes fungi play in providing bees withnutrients. Interestingly, Fomes fomentarius is a known endophyte ofbirch trees—meaning that they are part of the tree's natural immunesystem. The inventor hypothesizes that many of these endophytic fungiconfer antiviral properties on plants and bees—if encountered within adiscrete, diluted window, as well as other insects, as they forage ornest in wood hosting these fungi. But, if encountered in their pureform, many of these may, in fact, be toxic. Here is where humanintervention can help evolve a bridge of antiviral benefits otherwiseunlikely encountered in nature. The inventor believes theinter-relational dimensions wherein the biology of bees, insects, fungi,birds, bears and decomposing trees and plants all intersect will becomea fertile area of scientific research for helping and evolvingecosystems for decades to come.

Fomes fomentarius extracts dually reduce the Tobacco Mosaic Virus, aplant disease virus, and the Deformed Wing Virus, a virus harming bees.This inventor envisions making cocktails, complex mixtures of extractsfrom fungi, particularly wood decomposing polyporus fungi, to stave offmany viruses vectored by biting insects who transmit disease virusesharming plants. Finding a virus that dually harms plants and animals isexceedingly rare, if not yet unprecedented. Equally surprising is thatone extract from, for instance, Fomes fomentarius, would reduce bothbee-infecting and plant infecting viruses, and gives an enormousadvantage to agriculture. This inventor envisions many uniquecombinations of using extracts that reduce or prevent viruses that harmbees with extracts that reduce or prevent viruses that harm plants. Bygiving insects these cocktails, through direct feeding or indirectexposure, could reduce viruses transmitted by biting insects, orairborne viruses, saving agriculture many billions of dollars.

Of course, bears are not the only way to spread to trees Fomitopsis andother fungi that may improve bee heath. Any activity resulting increating wounds in trees, or in creating dead wood, creates a potentialfungal platform of bee benefit. The human use of woodchips as ‘beautybark’ or for making trails, or as a top dressing around ornamentals,would also serve to create a mycelial platform of benefit to bees.Ultimately, this means we can grow the mycelium of these fungi, enmasse, in a pre-sporulating or pre-conidial state, make mycelial‘landing pads’ for bees, or make extracts, and in doing so creating anew generation of bee attractants and nutrition customized accordingly.

The mycelium in nature acts as a cellular scaffolding in which and uponwhich millions of microbes exist, with many being harmful to themycelium. Pure culture mycelium in a laboratory, when healthy, hascomparatively very few (less than by many orders of magnitude) bacteriaand some of these bacteria help mycelium defend itself, only takingadvantage postmortem. These co-cultured mycelially resident bacteria aretypically not evident until the mycelium has been killed. These restingbacteria also produce antiviral and immunomodulating effects through theexpression of unique molecules, especially enzymes and lipids. Some ofthese bacteria can enhance resistance of the mycelium to outsidediseases. The beneficial bacteria associated with or in concert withmycelium that can help fight viruses and enhance host defense immunity,including but not limited to, species in the genera, sensu lato, ofPseudomonas, Bacillus, Acidophilus and Bifidobacterium. Moreover, thisconsortium can be highly inhibitory against Candida yeasts and bacterialpathogens in the genera, sensu lato, Serratia, Clostridium, Klebsiella,Bacteroides. Mycobacterium, Borrelia as well as other pathogens, fungal,bacterial, viral, or protozoal.

Another embodiment of this invention is to ferment the mycelium ofmedicinal mushrooms with Bifidobacterium bifidum, Lactobacillusplantarum, Lactobacillus acidophilus, Lactobacillus sakei, Leuconostoclactis, Streptococcus thermophiles and bacteriophages to make consumableand environmentally applicable compositions beneficial to the microbiomeof bees, animals (people).

Recent unpublished research funded by the inventor uses state-of-the-artNext-Generation (“NextGen”) sequencing to show that the consortium ofbacterial species selected by Stropharia rugosoannulata mycelium onfermented woodchips is several orders of separation, taxonomically, fromthe associated bacterial species of, for instance, Irpex lacteus. BothStropharia rugosoannulata and Irpex lacteus were inoculated intoseparate containers holding the same fermented woodchips. The testsproved the mycelium of mushroom species influences the subsequentlyevolving bacterial genome that is in close contact with the mycelialmycosphere (myco-rhizosphere), selecting subsets of mixed bacterialpopulations, and yet the mycelium growth rate, form and tenacity appearsextraordinarily healthy and vigorous for both fungal species. As such,this inventor anticipates that the microbiome—or mycobiome, i.e. themixed matrix of fungally selected bacteria—will produce healthy mycelialmats productive of sporulating fruitbodies but whose bacterially endowedmycelium is also friendly to bees and will also provide a bacterialcomponent which confers anti-pathogen resistance to, for instance,invading Nosema, a fungal microsporidium. This idiosyncratic consortiumof fungi and bacteria offers yet another complex bioshield of defense,protecting all the partners who depend upon each other—bees, mycelium,bacteria, plants, and animals, including humans. Fungal-bacterialmixtures can be customized to best benefit bees via a wide variety ofcompositions and methods of producing new bee supporting products.

Additionally, the antiviral effects seen in the fungal extracts arelikely to be partially attributable to the co-occurring residentbacteria that are endemic to fungal mycelial matrices, which alsoproject antiviral enzymes, antibiotics, and antiviral molecules. Theresults of antiviral activity from living mycelium may have enhancedantiviral effects due to the association of bacteria and fungi livingtogether, and their viromes, as a complex community and hence areextracted together in common.

The mycelium of Stropharia rugosoannulata, Fomitopsis officinalis,Fomitopsis pinicola, Piptoporus betulinus, Schizophyllum commune,Trametes elegans, Trametes versicolor species and many polyporoid andgilled basidiomycetes produce bioflavonoids, phenols and polyphenols,including coumarins and coumaric acids (both trans- and cis-o- andp-coumaric acids) which up-regulate genes in bees which code forcytochrome P450 enzymes as well as other enzymes critical for digestion,metabolism and toxin destruction. The effect of these mycelialcomponents such as coumarins, p-coumaric acid, o-coumaric acid or theirglycosides, is that they turn on more genes within bees which allow forthe bees to detoxify a wide range of toxins, particularly insecticides,miticides, herbicides, fungicides and pesticides, and augment the bee'sinnate immunity.

p-coumaric acid, found in both grains and lignin, is a monomer ofsporopollenin, the principal constituent of pollen cell walls andpropolis, the resinous compounds gathered and processed by bees to linewax cells. p-coumaric acid production is closely interrelated to theexpression of laccases in wood-rotting fungi. Laccase is a cellulaseenzyme that breaks down lignin in wood, creating derivative compoundspalatable to insects as food, as well as creating habitats (bees cantake up residence in tunnels bored by mycophagous beetles). As fungi rotwood, breaking down lignin, they also weep water, rich in thesep-coumaric and nutraceutical compounds beneficial to bees. The morelaccases, the more p-coumaric acid expressed by the mycelium, and viceversa, accelerating decomposition. The more the wood rots, the morefungal polysaccharides (sugars) and ultimately the more these compoundswill be in the fungal exudates that the bees seek and from which theybenefit. That wood rotting fungi produce p-coumaric acids and thatcoumarins can be bio-converted into p-coumaric acids is yet anotheradvantage of this invention.

As was noted by Terrón et al., structurally closely related aromaticcompounds have different effects on laccase activity and on Icc geneexpression in the ligninolytic fungus Trametes sp. 1-62, Fungal Genet.Biol., October 2004; 41(10):954-62: “Nine phenolic compounds (p-coumaricacid, ferulic acid, guaiacol, syringol, p-methoxyphenol, pyrocatechol,phloroglucinol, 3,5-dihydroxybenzoic acid, and syringaldazine) weretested for their ability to increase laccase production in theligninolytic basidiomycete Trametes sp. 1-62. All these compoundsresulted in increases in laccase activity, with the highest levels beingdetected in the presence of p-coumaric acid (273-fold) and guaiacol(73-fold).”

Interestingly, many of the grains preferred for mycelial spawnproduction for mushroom industry (see Growing Gourmet & MedicinalMushrooms by the inventor, Paul Stamets, 1993, 2000, Ten Speed Press,Berkeley) are also rich sources of p-coumaric acids and may be useful inbee attractant compositions. The primary phenolic acids in rice grainwere identified as p-coumaric acid, ferulic acid, and sinapinic acid.

P-coumaric acid is not only in the grains preferred for mushroom spawnproduction but they are also generated during the normal life cycle ofmushrooms, especially prior to primordia formation. P-coumaric acid is apotent inhibitor of tyrosinase, the enzyme essential for melaninization.The presence and abundance of p-coumaric acid interferes with theproduction of darkly colored pigments. Ultraviolet light stimulates thephotodecomposition of p-coumaric acids, enabling melanization andtriggering primordia formation. Once primordia forms, p-coumaric acidsdegrade into p-hydroxybenzoic acid. (Sachan et al., Transformingp-coumaric acid into p-hydroxybenzoic acid by the mycelial culture of awhite rot fungus Schizophyllum commune, 2010, African Journal ofMicrobiology 4:267-273. ) As an example, but not one of limitation, themycelium of Auricularia auricula (A. auricularia-judae), when grown inculture is whitish and lacks melanin but contains p-coumaric acids. Whenthe mushroom mycelium is exposed to light, the mycelium bio-transformsto create dark brown fruitbodies, which are higher in melanin as theymature, with p-coumaric acids, an inhibitor of melanin, concurrentlydeclining. This is one example and a strong argument for the benefit ofusing lightly colored mycelium, pre-melaninization as a source ofmycelium for making extracts beneficial to bees due to its innatep-coumaric acid content compounded by the native content of p-coumaricacids in the grains that are used for spawn production for growingmycelium. Interestingly, the ideal interface for capturing the bestbenefits from mycelium for its nutraceutical and p-coumaric acidcontents, is a short window, often of just a few days in length, beforeand directly after light exposure, but before dark colored fruitbodydevelopment beyond the white primordial stage ensues. UV Lightstimulates vitamin D pathways as well as activates tyrosinaseproduction, which leads to pigmentation, at a transitional stage in thelife cycle corresponding to decreases in laccases and p-coumaric acids.

Given that some of the most abundant laccase producers yet tested thusfar are Ganoderma lucidum, Trametes versicolor and Pleurotus ostreatus,these species are specifically preferred for use in creatingbee-beneficial mixtures.

“We seek to understand the botanical sources and biological activitiesof resins in the field and how resin foraging behavior changes inresponse to environmental factors, such as infection and otherbiological stresses. If we can discover plants with preferable and moreantimicrobial resins in different regions, it should be possible tobetter create environments that promote bee health by supportingbehaviors and managerial strategies that lead to natural diseaseresistance.” (Wilson et al., Metabolomics reveals the origins ofantimicrobial resins collected by honey bees. PLoS One 8(10): e77512,page 11. ) The present inventor suggests that fungi and fungal mycelium,including fungal attractants, fungal entomopathogens, fungalimmunostimulators, fungal antivirals, antibacterials and antifungals cansimilarly support bee health and lead to natural resistance to diseasesand pesticides. Ecosystems and economies benefit from bees that wouldotherwise suffer without these myco-remedies.

This inventor also anticipates that pollinating insects and animals(bats) will also benefit from the effects of this invention. It is alsoexpected that birds may similarly benefit from similar integrated fungalsolutions via addition to nectar feeders and bird foods through the upregulation of immunological and detoxification genes as well asreceiving antiviral benefits, thus extending longevity.

This invention enables the creation of ‘smart foods’ or ‘smartnutraceuticals’ that can help prevent neuropathy by stimulatingneurogenesis. Filamentous, basidiomycetous fungi are sources ofneuroregenerative compounds. Species of Hericium (including but notlimited to Hericium erinaceus, Hericium coralloides and Hericiumabietis) produce potent nerve growth factors causing regeneration ofmyelin on the axons of nerves and nerve regeneration. See Stamets,Lion's Mane: A Mushroom That Improves Your Memory and Mood?, The Blog,Huffington Post Healthy Living, Aug. 8 2012.

Psilocybin and psilocybin-producing fungi, including but not limited tospecies of Psilocybe, Panaeolus, Gymnopilus, Pluteus and Conocybe suchas Psilocybe azurescens, Psilocybe cyanescens, Psilocybe allenii,Psilocybe cyanofibrillosa, Psilocybe cubensis, Psilocybeovoideocystidiata, Psilocybe subaeruginosa, Copelandia Panaeolus(Copelandia cyanescens, Copelandia tropicalis, Copelandia bispora),Pluteus salicinus, Gymnopilus luteofolius, Gymnopilus spectabilis,Conocybe cyanopus and Conocybe smithii can trigger neurogenesis. (SeeCatlow et al., Effects of psilocybin on hippocampal neurogenesis andextinction of trace fear conditioning, Exp Brain Res (2013) 228:481-491DOI 10.1007/s00221-013-3579-0). Individually or in combination, mixturesof extracts of psilocybin mushroom and Hericium mushroom fruitbodies, ormore preferably their mycelial extracts, could help repair neuronsdamaged by toxins, cholinergic pesticides, fungicides, herbicides,glyphosates, oxidation, old age, or other sources of neuro-damagingtoxins. The net effect of ingesting these mixtures of nerve regeneratingHericium and psilocybin species would improve the neurological health ofbees through neurogenesis and re-myelination, and indeed of animals,including humans. Another, improved form of “smart mycohoney” mightincorporate these elements for the benefits of bees and people,improving cognition, preventing or repairing neuropathies presentingthemselves as diseases to humans within scope of the definitions forAlzheimer's, Parkinson's, Parkinsonisms, MS (multiple sclerosis), or asyet uncategorized forms of neurological impairment. Indeed, suchcombinations could increase intelligence, sensory abilities, memory,reflexes, reaction times, and problem-solving abilities. Moreover, tothe above mixture, vitamins can be added for further enhancement ofbeneficial properties. The addition of vitamin D—either from UV exposedfungal cells or from external sources, with or without Vitamin B(niacin, nicotinic acid, or related congener), enhance neurogenesis andare preferred ingredients. As such a smart nutraceutical in many formsare possible, including a ‘smart mycohoney’ or ‘smart mycosyrup’ both ofwhich are anticipated to be within the scope of this invention. Such amycosyrup can be reduced into solid or powdered form added to any foodconsumed by animals, or by any means known to pharmaceutical science.

Example 40

The use of these antiviral fungal extracts combined with the rapidlyevolving CRISPR technology will be led to new breakthroughs inpotentiating the antiviral and virostatic effects in treating animals,plants, and bacteria. Moreover, the author anticipates that when usingthese antiviral and immune supporting fungal mycelium and extracts,competitive, non-threatening or “beneficial” viruses can be favorablyselected from the virome. Such selection may require multiple dosingregimens of different fungal species and fungal-bacterial communities sothat a more potent mixture can be better tuned within the virome, makingthe virome better populated with advantageous viruses and the cells thathost them. These selected subset populations of beneficial viruses willcompete with pathogenic viruses within cells, diminishing theirdeleterious effects of pathogenic viruses. Moreover, these extracts andtheir fractions, will further adjust the bacterial and fungalmicrobiomes to the benefit of the overall health and longevity of thehosts, including but not limited to organisms that are virallysusceptible, such as bees, animals, plants, and bacteria. By addingCRSIPR technologies for inserting genes into and from bacteria andviruses, and with the use of selectively active antiviral extracts madefrom fungal mycelia, the microbiomes and viromes extending thelongevity, strength, health and disease resistance of the infected orto-be-infected organism at risk can be tuned for the benefit offortifying health and survival.

Example 41

By adding preconidial mycelium of Metarhizium anisopliae or Metarhiziumbrunneum in any form (dried, freeze dried, fresh) to seeds, theMetarhizium mycelium can enter within the plant tissues, postgermination, and confer to the plant a mycoinsecticidal defense againstpredaceous insects and arthropods. This method and composition can bepresented into wildflower seeds, or to other seeds, where the plantsproduce flowers which bees frequent, which will help them better defendthemselves against, for example, Varrroa mites, from contact with theplant or the pollen. Moreover, the use of Metarhizium in direct contactwith seeds which results in invasion of the Metarhizium mycelium intothe plants can benefit many plants against many insect and arthropodpredators. This has wide applications for benefitting agricultureworldwide.

Example 42

This invention can also be instrumental for terraforming in the creationof new life-sustaining habitats, including on other planets. Knowing thecomplex interrelationships that are necessary to achieve aself-propelling, yet balanced ecosystem requires a level of biodiversitythat currently is beyond our best practices or knowledge. Thisinvention—fortifying the immune system and lifespan of bees—as well asits animal and plant inhabitants—is a strong underpinning of thebiological foundation necessary for designing and implementingterraforming on other planets, or even repairing habitats currentlydamaged on Earth. Mycoterraforming can include creating uniquecompositions consisting of saprophytic, endophytic, and mycorrhizalfungi, paired for benefitting the long-term health of constructedhabitats.

For instance, utilizing the paper and cellulose products containinggoods sent along with colonists to Mars, and to help recycle humanwaste, spores of saprophytic Pleurotus, Stropharia, Coprinus andPsilocybe species can be sent to decompose this waste. EndomycorrhizalGlomus species and ectomycorrhizal Pisolithus fungi can be combined withthe endophytic Curvularia, Aspergillus, Fusarium, Penicillium,Gilmaniella, and Arthrobotrys fungi, to help jumpstart ecosystems. Sinceseed production will be crucial for expanding terraforming and sincemany if not most planets benefit from buzz pollination or directpollination by insects, including but not limited to honey bees, theextracts described herein can be desiccated into a form useful fortransport, frozen, and thawed when needed. Once sufficient biomass istransformed into soil that can sustain woody plants and trees, thesurvival of bees in particular, will be critical for habitat evolution.After this mycologically derived soil has been created, this author hasfound that many bee-benefitting fungi produce mycelia from digestingcellulosic substrates and, in the course of decomposition, producesanti-viral and longevity enhancing exudates than can be made intoextracts.

Taking bees into space creates obvious problems to the layperson, butentomologists and even amateur beekeepers are more circumspect, knowinghow to co-exist with bees in mutually beneficial relationships, withoutthe fear of being stung arresting their activities. Specific concoctionscan be designed by astro-entomologists and astro-mycologists, in concertwith other biologists and bioengineers, to kickstart ecosystems that canquickly emerge into life-sustaining habitats, allowing for human oranimal colonization on other planets. As such designing, maintaining,and improving biomes—microbiomes, mycobiomes, and viromes—areanticipated to take advantage of this inventor's ideas for sustainablespace travel and for terraforming other planets.

Moreover, these genomes may be pre-programmed to encode the vastness ofhuman knowledge, bioaccumulating more knowledge, as these livingmycobased systems emerge, adapt, learn, and become wells of wisdom.Habitats then can become living libraries that form the body intellectfrom which humans can benefit as we travel into the cosmos.

What is claimed:
 1. A composition for improving bee health comprisingeffective amounts of: 1% or less by volume of one or more aqueousethanol mycelial extracts of Ganoderma applanatum, Ganoderma resinaceum,Fomes fomentarius, Fomitopsis officinalis, Trametes versicolor, orcombinations thereof; one or more bee feeding supplements; and one ormore preservatives.
 2. The composition of claim 1, wherein the mycelialextract is from Ganoderma applanatum.
 3. The composition of claim 1,wherein the mycelial extract is from Ganoderma resinaceum.
 4. Thecomposition of claim 1, wherein the mycelial extract is from Fomesfomentarius.
 5. The composition of claim 1, wherein the mycelial extractis from Fomitopsis officinalis.
 6. The composition of claim 1, whereinthe mycelial extract is from Trametes versicolor.
 7. The composition ofclaim 1, wherein the composition further comprises a mycelial extractfrom Antrodia cinnamonea, Fomitopsis pinicola, Ganoderma atrum,Ganoderma brownii, Ganoderma curtisii, Ganoderma lucidum, Ganodermalingzhi, Ganoderma oregonense, Ganoderma tsugae, Fomitiporia robusta,Heterobasidion annosum, Inonotus hispidus, Inonotus andersonii, Inonotusdryadeus, Inonotus obliquus, Laetiporus cincinnatus, Laetiporussulphureus, Laetiporus conifericola, Lenzites betulina, Phellinusigniarius, Phellinus linteus, Phellinus pini, Piptoporus betulinus,Polyporus elegans, Stereum complicatum, Stereum hirsutum, Stereumostrea, Schizophyllum commune, Trametes elegans, Trametes gibbosa,Trametes hirsuta, Trametes villosa, Trametes cingulata, Trametesochracea, Trametes pubescens, Trametes ectypa, Trametes aesculi,Wolfiporia cocos, Agaricus augustus, Agaricus blazei, Agaricus bonardii,Agaricus brasiliensis, Agaricus campestris, Agaricus lilaceps, Agaricussubrufescens, Agaricus sylvicola, Agrocybe pediades, Agrocybe aegerita,Agrocybe arvalis, Agrocybe praecox, Clitocybe odora, Conocybe cyanopus,Conocybe lacteus, Conocybe rickenii, Conocybe smithii, Conocybe tenera,Coprinopsis nivea, Coprinopsis lagopus, Coprinus comatus, Coprinusmicaceus, Gymnopus hydrophilus, Gymnopus peronatus, Hypholoma aurantiaca(Leratiomyces ceres), Hypholoma capnoides, Hypholoma sublateritium,Hypsizygus marmoreus, Hypsizygus tessulatus, Hypsizygus ulmarius,Lentinus ponderosus, Lepiota procera (Macrolepiota procera), Lepiotarachodes (Chlorophyllum rachodes), Lepista nuda, Mycena alcalina, Mycenapura, Mycena aurantiidisca, Panellus serotinus, Panaeolus foenisecii,Panaeolus subbalteatus, Pleurotus columbinus, Pleurotus ostreatus,Pleurotus cystidiosus, Pleurotus pulmonarius, Pleurotus sapidus,Pleurotus tuberregium, Panellus stipticus, Panellus serotinus, Pluteuscervinus, Psathyrella aquatica, Psathyrella candolleana, Psathyrellahydrophila, Psilocybe allenii, Psilocybe azurescens, Psilocybecaerulescens, Psilocybe coprophila, Psilocybe cubensis, Psilocybecyanescens, Psilocybe ovoideocystidiata, Psilocybe stuntzii, Psilocybesubaeruginosa, Stropharia aeruginosa, Stropharia cyanea, Strophariarugosoannulata, Stropharia semiglobata, Stropharia semigloboides,Stropharia squamosa, Stropharia thrausta, Stropharia umbonatescens,Termitomyces robusta, Volvariella bombycina, Volvariella volvacea, orcombinations thereof; and wherein the composition comprises a totaleffective amount of about 1% or less by volume of mycelium extract. 8.The composition of claim 1, wherein the bee feeding supplements compriseone or more of water, sugars, sugar syrup, high fructose corn syrupwater, bee candy, nectar, pollen, pollen patties, grease patties,propolis, bees wax, bee sprays, bee feed, protein supplements, orcombinations thereof.
 9. The composition of claim 1, wherein thepreservative comprises ethanol, isopropanol, methanol, butyl alcohol,other C₂-C₆ alcohols, benzalkonium chloride, benzalkonium chloridesolution, benzethonium chloride, benzoic acid, benzyl alcohol,butylparaben, cetylpyridinium chloride, chlorobutanol, chlorocresol,cresol, dehydroacetic acid, ethylparaben, methylparaben, methylparabensodium, phenol, phenylethyl alcohol, phenylmercuric acetate,phenylmercuric nitrate, potassium benzoate, potassium sorbate,propylparaben, propylparaben sodium, sodium benzoate, sodiumdehydroacetate, sodium propionate, sorbic acid, thimerosal, thymol, orcombinations thereof.
 10. The composition of claim 1, wherein theextract comprises an aqueous ethanol mycelium extract; a dried aqueousethanol mycelium extract; a supernatant remaining after precipitation ofan aqueous mycelium extract with ethanol; a supernatant from an aqueousethanol mycelium extract having a portion of solvent removed; asupernatant from aqueous ethanol mycelium extract having solventremoved; a supernatant from an aqueous ethanol mycelium extract having aportion of solvent and all precipitate removed; a supernatant fromaqueous ethanol mycelium extract having both solvent and precipitateremoved; steam distilled ethanol extracts; microwave-assisted ethanolextracts; or combinations thereof.
 11. The composition of claim 1,wherein the mycelium is cultivated on a substrate comprising solidsubstrates or liquid substrates.
 12. The composition of claim 1, whereinthe mycelium is cultivated on a substrate comprising deciduous orconiferous sawdust.
 13. The composition of claim 1, wherein the myceliumis cultivated on a substrate comprising birch or alder sawdust.
 14. Thecomposition of claim 1, wherein the composition improves bee health byincreasing longevity by more than about 1%.
 15. The composition of claim1, wherein the composition improves bee health by increasing longevityby more than about 3%.
 16. The composition of claim 1, wherein thecomposition improves bee health by increasing longevity by more thanabout 5%.
 17. The composition of claim 1, wherein the compositionimproves bee health by reducing viral load by more than about 1%. 18.The composition of claim 1, wherein the composition improves bee healthby reducing viral load by more than about 15%.
 19. The composition ofclaim 1, wherein the composition improves bee health by reducing viralload by more than about 25%.
 20. The composition of claim 1, wherein thecomposition improves bee health by increasing longevity and reducingviral load by an LV index of more about than
 1. 21. The composition ofclaim 1, wherein the composition improves bee health by increasinglongevity and reducing viral load by an LV index of more than about 50.22. The composition of claim 1, wherein the composition improves beehealth by increasing longevity and reducing viral load by an LV index ofmore than about 200.